Secondary Reagents. Secondary Reagents. biolegend.com 551

Transcription

1 Anti-Hamster IgG (Armenian) Anti-Hamster IgG (Syrian) Anti-Human Ig light chain κ Anti-Human Ig light chain λ Anti-Human IgD Anti-Human IgE Anti-Human IgG Fc Anti-Human IgM Anti-Mouse IgA Anti-Mouse IgD Anti-Mouse IgD a (Igh-5a) Anti-Mouse IgE Anti-Mouse IgE a Anti-Mouse IgE b Anti-Mouse IgG Anti-Mouse IgG1 (Goat) Anti-Mouse IgG2a Anti-Mouse IgG2a (Goat) Anti-Mouse IgG2b Anti-Mouse IgG Anti-Mouse IgM Anti-Mouse IgM a (Igh-6a) Anti-Mouse IgM b (Igh-6b) Anti-Mouse Ig light chain κ Anti-Mouse Ig light chain λ Anti-Mouse IgG Anti-Rabbit IgG Anti-Rat IgG Anti-Rat IgG2a Anti-Rat IgG2b Anti-Rat IgG2c Anti-Rat IgM Anti-Rat Ig light chain κ Anti-Rat Ig light chain λ Anti-Rat IgG Avidin, Avidin, HRP , AKP , , /Cy , Brilliant Violet , Cy , Cy , DyLight , DyLight , DyLight , DyLight , , HRP , , /Cy , /Cy , PerCP , PerCP/Cy Anti-Allophycocyanin Anti Anti-Fluorescein () Anti-KLH Anti-Phycoerythrin () biolegend.com cs@biolegend.com 551

2 Anti-Hamster IgG (Armenian) Poly4055 Isotype: Goat polyclonal Ig Reactivity: Hamster (minimal x-reactivity) FC or ELISA (depending on format) Quality tested IF, WB, IHC Reported in the literature This polyclonal antibody reacts with the heavy chains of IgG and the light chains common to most immunoglobulins from Armenian hamster. No cross-reactivity has been detected against non-immunoglobulin serum proteins. This antibody has been solid-phase absorbed to ensure minimal cross-reaction with mouse, rat, human, bovine, and rabbit immunoglobulins, but may cross-react with Armenian hamster IgM and Syrian hamster immunoglobulins. Poly $140 DyLight $140 DyLight DyLight Anti-Hamster IgG (Syrian) Poly4056 Isotype: Goat polyclonal Ig Reactivity: Hamster (minimal x-reactivity) ELISA, IF, WB, IHC Reported in the literature This polyclonal antibody reacts with the heavy chains of IgG and the light chains common to most immunoglobulins from Syrian hamster. No cross-reactivity has been detected against non- immunoglobulin serum proteins. This antibody has been solid-phase absorbed to ensure minimal cross-reaction with mouse, rat, human, bovine, and rabbit immunoglobulins, but may cross-react with Syrian hamster IgM and Armenian hamster immunoglobulins. Poly $ $140 Anti-Human Ig light chain κ Clone: MHK-49 Isotype: Mouse IgG1, κ Immunogen: Human IgGs and IgM, IgE, IgA cocktail Reactivity: Human anti-κ (MHK-49) anti-λ (MHL-38) CD19+ lymphocytes stained with anti-κ (MHK-49) and anti-λ (MHL-38) immunofluorescent staining or ELISA analysis. Clone MHK $ $ $ $ $245 FC, IC ELISA Reported in the literature The MHK-49 antibody reacts with both soluble and membrane human immunoglobulin light chain kappa (k). It does not react with human immunoglobulin light chain lambda (λ) or heavy chain (μ). The MHK-49 antibody can be used as primary or secondary reagent for $160 Alexa Fluor Anti-Human Ig light chain λ Clone: MHL-38 Isotype: Mouse IgG2a, κ Immunogen: Human IgGs and IgM, IgE, IgA cocktail Reactivity: Human anti-κ (MHK-49) anti-λ (MHL-38) CD19+ lymphocytes stained with anti-κ (MHK-49) and anti-λ (MHL-38) The MHL-38 antibody reacts with both soluble and membrane human immunoglobulin light chain lambda (λ). It does not react with human immunoglobulin light chain kappa (k) or heavy chain (μ). The MHL-38 antibody can be used as primary or secondary reagent for immunofluorescent staining or ELISA analysis. Clone MHL $ $ $ $160 Alexa Fluor 488 Alexa Fluor Anti-Human IgD FC, IC ELISA Reported in the literature Clone: IA6-2 Isotype: Mouse IgG2a, κ Immunogen: Human IgD Reactivity: Human $200 Pacific Blue $ $ Toll Free BIOLEGEND ( )

3 IA6-2 CD19 (HIB19) Alexa Flour 647 Human peripheral blood lymphocytes stained with IA6-2 and CD19 (HIB19) Alexa Fluor 647 IgD is a member of the Immunoglobulin (Ig) family, is expressed in naïve B cells, has 3 Ig-like domains, and exists in a transmembrane and a soluble form. In general, IgD is not secreted and usually is not expressed after the Ig isotype switch. After antigen binding, IgD signals trough the CD79a/CD79b (Igα/Igβ) heterodimer, resulting in the activation of the B cell. Application References: 1. Chen, K., et al Nat. Immunol. 10: Lee, C.H., et al J. Exp. Med. 203: Sutter, J.A., et al Clin. Immunol. 126:282. Clone IA $ $ $135 /Cy7 Anti-Human IgE $ $ $215 PerCP/Cy $ $325 Clone: MHE-18 Isotype: Mouse IgG1, κ Immunogen: Human IgGs and IgM, IgE, IgA cocktail Reactivity: Human The MHE-18 antibody reacts with both soluble and membrane human immunoglobulin E (IgE). It does not react with other Ig isotypes. IgE is the seventh immunoglobulin made by B cells in the immune response. The MHE-18 antibody can be used as primary or secondary reagent for immunofluorescent staining or ELISA analysis. ELISA, IF, IHC Reported in the literature MHE-18 CD203c Overnight cultured human PBMCs stained with anti-human IgE (MHE-18) and biotinylated CD203c, followed by Sav- (gated on lymphocyte population) Anti-Human IgG Fc Clone MHE $ $ $130 Clone: HP6017 Isotype: Mouse IgG2a, κ Immunogen: human immunoglobulin Reactivity: Human $285 Human DLL4 transfected CHO cells were incubated with (top) or without (bottom) recombinant human Notch 1-IgG Fc fusion protein and then stained with human IgG Fc (clone HP6017) (filled histogram) or mouse IgG2a, k isotype control (open histogram) IgG Fc is a homodimer composed by the constant region of the two heavy chains that conform the IgG molecule. The Fc fragment mediates opsonisation, antibody dependent cellular cytotoxicity (ADCC) and complement activation, through binding to Fc receptors such as CD16, CD32, CD64, and the complement factor C1. Application References: 1. Reime, C.B., et al Hybridoma 3: Jefferis, R., et al Immunol. Lett. 10:223. Clone HP $ $ $110 Anti-Human IgM $265 Clone: MHM-88 Isotype: Mouse IgG1, κ Immunogen: Human IgGs and IgM, IgE, IgA cocktail Reactivity: Human FC, IC ELISA Reported in the literature CD19 MHM-88 Human peripheral blood lymphocytes stained with CD19 (HIB19) and MHM-88 The MHM-88 antibody reacts with both soluble and membrane human immunoglobulin M (IgM). It does not react with other Ig isotypes. IgM is the first immunoglobulin made by B cells in the immune response. Surface IgM is expressed on immature and mature B cells, while IgM heavy (μ) chain is expressed intracellularly in pre-b cells. The MHM-88 antibody can be used as primary or secondary reagent for immunofluorescent staining or ELISA analysis. Clone MHM $ $200 Pacific Blue $ $ $ $ $245 PerCP/Cy $ $ $140 /Cy7 Brilliant Violet $ $295 biolegend.com cs@biolegend.com 553

4 Anti-Mouse IgA Clone: RMA-1 Isotype: Rat IgG ELISA, FC The RMA-1 monoclonal antibody reacts with immunoglobulin A (IgA) in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. The RMA-1 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone RMA Anti-Mouse IgD Clone: 11-26c.2a C57BL/6 mouse splenocytes stained with 11-26c.2a Isotype: Rat IgG2a, κ FC 1,4,5 Quality tested IHC (Frozen) 2,3 Reported in the literature The 11-26c.2a antibody reacts with immunoglobulin D in all tested mouse haplotypes. The antibody binds membrane IgD expressed on most B cells. Surface IgD is an important B cell differentiation marker. The 11-26c.2a antibody neither induces proliferation of splenic B cells nor induces B cell activation. Application References: 1. Nitschke, L., et al PNAS 90: Weih, D., et al J. Immunol. 167: Koni, P.A., et al J. Exp. Med. 193: Ahuja, A., et al J. Immunol. 179: Haynes, N.M., et al J. Immunol. 179: Clone 11-26c.2a $ $ $ $220 Alexa Fluor $ $110 /Cy $ $275 Pacific Blue $ $240 Alexa Fluor PerCP/Cy Anti-Mouse IgD a (Igh-5a) $295 Other Name: Igh-5.4 Clone: AMS-9.1 Isotype: Mouse (SJL) IgG2b, κ BALB/c splenocytes stained with AMS-9.1 Pacific Blue IHC (Frozen) Reported in the literature This antibody was raised by immunization with BALB/c mouse splenocytes. It specifically reacts with mouse IgD of Igh-Ca haplotype, but does not react with b and c haplotypes. This antibody is able to induce B cell activation and proliferation. Application References: 1. Stall, AM., et al J. Immunol. 132: Goroff, D.K., et al J. Immunol. 136: Clone AMS-9.1 Pacific Blue $ $80 Anti-Mouse IgE $ $195 Clone: RME-1 Isotype: Rat IgG1, κ FC or ELISA (depending on format) Quality tested The RME-1 monoclonal antibody reacts with immunoglobulin E (IgE) in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. The RME-1 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone RME Optical Density (450nm) $ $ $ $ Mouse IgE ELISA MAX Set Standard Set Contents: 1. Mouse IgE-specific pre-titrated Capture Antibody 2. Mouse IgE-specific pre-titrated ylated Detection Antibody 3. Mouse IgE Standard 4. Avidin-Horseradish Peroxidase Mouse IgE (pg/ml) Deluxe Set Contents: All reagents (1-4) in the Standard Set, plus: 5. TMB Substrate Reagent 6. Coating Buffer (5 ) 7. Assay Diluent (5X) 8. NUNC MaxiSorp 96-well ELISA plates (uncoated) Application References: 1. Wang, W., et al J. Immunol. 178: Toll Free BIOLEGEND ( )

5 Mouse IgE ELISA MAX Set Standard Sets Plates Plates $ Plates $550 Anti-Mouse IgE a Plates $295 Deluxe Sets Plates $ Plates $665 The UH297 antibody reacts specifically with mouse soluble immunoglobulin E of Igh-Ca haplotype (e.g., A, AKR, BALB/c, C3H/He, CBA). It does not react with IgE Igh-cb haplotype (e.g., C57BL/6, C57BL/10, SJL) and other Ig isotypes. UH297 mab can be used for immunofluorescent staining of cell surface IgE a and ELISA applications. Clone: UH297 Isotype: Rat IgG1, κ Immunogen: BALB/c mouse IgE anti-dnp mab from DNPε 117 hybridoma FC, WB Reported in the literature Application References: 1. Usui, M. et al Immunogenetics. 37: Yamada, M. et al Immunogenetics. 40: Clone UH $215 Anti-Mouse IgE b The JKS-6 antibody reacts with mouse soluble and cell surface immunoglobulin E (IgE) of Igh-C b haplotype (e.g., C57BL/6, C57BL/10, SJL). It does not react with IgE Igh-c a haplotype(e.g., A, AKR, BALB/c, C3H/He, CBA) and any other Ig isotypes. JKS-6 mab can be used for immunofluorescent staining of cell surface IgE b and ELISA application. Clone: JKS-6 Isotype: Rat IgG2a, κ FC Reported in the literature Application References: 1. Yamada, M. et al Immunogenetics. 40: Clone JKS $215 Anti-Mouse IgG1 Clone: RMG1-1 Isotype: Rat IgG FC or ELISA (depending on format) Quality tested IHC, IF Reported in the literature The RMG1-1 monoclonal antibody reacts with immunoglobulin G1 (IgG1) in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. The RMG1-1 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Application References: 1. Rucci, F., et al P. Nat. Acad. Sci. USA 107:3024 Clone RMG $ $ Anti-Mouse IgG1 (Goat) Poly24091 Isotype: Goat polyclonal Ig FC, IF DyLight -conjugated goat anti-mouse IgG1 reacts with immunoglobulin G1 (IgG1) in all tested mouse haplotypes (Igh-a and b). This antibody has been solid-phase absorbed to ensure Hela cells were stained with minimal cross-reaction with rat, anti-tubulin-α and secondarily human, bovine, horse, and rabbit immunoglobulins. DyLight - labeled with DyLight 594 goat anti-mouse IgG1 (Poly24091). conjugated goat anti-mouse IgG1 is useful for direct immunofluorescent staining or as a second step reagent for indirect immunofluorescent staining when used in conjunction with primary antibodies. Poly24091 DyLight DyLight Anti-Mouse IgG2a DyLight DyLight Clone: RMG2a-62 Isotype: Rat IgG FC or ELISA (depending on format) Quality tested IHC, IF Reported in the literature The RMG2a-62 monoclonal antibody reacts with immunoglobulin G2a (IgG2a) in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. The RMG2a-62 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Application References: 1. Gavin, A. L., et al Science 314: Clone RMG2a $ biolegend.com cs@biolegend.com 555

6 Anti-Mouse IgG2a (Goat) Anti-Mouse IgM Poly24092 FC, IF Isotype: Goat polyclonal Ig DyLight -conjugated goat anti-mouse IgG2a reacts with immunoglobulin G2a (IgG2a) in all tested mouse haplotypes (Igh-a and b). This antibody has been solid-phase absorbed to ensure minimal cross-reaction with rat, human, bovine, horse, and rabbit immunoglobulins. DyLight goat anti-mouse IgG2a is useful for direct immunofluorescent staining or as a second step reagent for indirect immunofluorescent staining when used in conjunction with primary antibodies. Poly24092 DyLight DyLight Anti-Mouse IgG2b DyLight DyLight DyLight Clone: RMG2b-1 Isotype: Rat IgG FC or ELISA (depending on format) Quality tested The RMG2b-1 monoclonal antibody reacts with immunoglobulin G2b (IgG2b) in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. The RMG2b-1 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone RMG2b Anti-Mouse IgG Clone: RMG3-1 Isotype: Rat IgG FC Reported in the literature The RMG3-1 monoclonal antibody reacts with immunoglobulin G3 (IgG3) in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. The RMG3-1 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Application References: 1. Fiorentini, C., et al Mol Pharmacol. 74:59. Clone RMG Clone: RMM-1 Isotype: Rat IgG2a, κ FC Quality tested ELISA Reported in the literature 6D5 (CD19) RMM-1 C57BL/6 mouse splenocytes stained with CD19 and RMM-1 The RMM-1 antibody reacts with both soluble and membrane immunoglobulin M in all tested mouse haplotypes (Igh-a and b). It does not react with other isotypes. IgM is the first immunoglobulin made by B cells in the immune response. Surface IgM is expressed on the majority of mature B cells. The RMM-1 antibody may be used as primary or secondary reagent for immunofluorescent staining or ELISA analysis. Clone RMM $ $85 /Cy $ $ $75 /Cy $ $195 PerCP/Cy $ $ $170 Brilliant Violet μl $160 Anti-Mouse IgM a (Igh-6a) Clone: MA-69 Isotype: Mouse (C57BL/6) IgG1, κ IHC (Frozen), ELISA Reported in the literature Balb/c mouse splenocytes stained with MA-69 This antibody was raised by immunization with IgM a mab TC31. It specifically reacts with mouse IgM of Igh-Ca and the related haplotypes (c, f, g, h, and j). It has been reported that beads coated with MA-69 antibody can stimulate B cell proliferation. Application References: 1. Sieckmann, D., et al Hybridoma 10: Stall, A.M., Weir s Handbook of Experimental Immunology pp. 27. Clone MA $ $ $ $ $ Toll Free BIOLEGEND ( )

7 Anti-Mouse IgM b (Igh-6b) Clone: AF6-78 C57BL/6 mouse splenocytes stained with AF6 78 ELISA Reported in the literature Isotype: Mouse (BALB/c) IgG1, κ BALB/c mouse splenocytes stained with AF6 78 This monoclonal antibody was raised by immunization with C57BL/10 mouse splenocytes. It specifically reacts with IgM of Igh-Cb and the related haplotypes (d and n), but not with the free μ heavy chain of IgM. Some cross-reaction with IgM of Igh-Ce can be detected with this antibody. Application References: 1. Stall, A.M., et al J. Immunol. 132: Velardi, A., et al J. Immunol. 133: Clone AF $ $ $ $290 C57BL/6 mouse splenocytes stained with RML-42 ELISA or immunofluorescent analysis. Clone RML Anti-Mouse IgG FC, ICFC, or ELISA (depending on format) Quality tested The RML-42 monoclonal antibody reacts with immunoglobulin light chain lambda in all tested mouse haplotypes (Igh-a and b). It does not react with kappa chain. The RML-42 monoclonal antibody may be used as primary or secondary reagent for $265 Poly4053 Isotype: Goat polyclonal Ig $265 Anti-Mouse Ig light chain κ Clone: RMK-12 Isotype: Rat IgG FC or ELISA (depending on format) Quality tested The RMK-12 monoclonal antibody reacts with immunoglobulin light chain kappa in all tested mouse haplotypes (Igh-a and b). It does not react with lambda chain. The RMK-12 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone RMK Anti-Mouse Ig light chain λ Clone: RML-42 Isotype: Rat IgG Human peripheral blood lymphocytes stained with purified mouse anti-human CD3 (UCHT1), followed antimouse IgG (Poly4053) /Cy7 FC or ELISA (depending on format) Quality tested IF, WB, IHC Reported in the literature 293E cells were stained with purified anti-tubulin-alpha (clone TU-01) and then BV421 anti-mouse IgG (Poly4053) (blue) as the secondary antibody. Nuclei were stained with propidium iodide (red). This polyclonal goat anti-mouse IgG antibody reacts with the heavy chains of mouse IgG and with the light chains common to most mouse immunoglobulins. No cross-reactivity has been detected against non-immunoglobulin serum proteins. This antibody has been solid-phase absorbed to ensure minimal crossreaction with rat, human, bovine, horse, and rabbit immunoglobulins, but it may cross-react with other subclasses of mouse immunoglobulins. Poly /Cy HRP μl $ $200 Cy /Cy $230 DyLight $180 PerCP/Cy DyLight Continues biolegend.com cs@biolegend.com 557

8 Anti-Mouse IgG continued DyLight Poly4060 DyLight AKP μl Isotype: Rat polyclonal IgG Brilliant Violet μl $185 This polyclonal rat anti-mouse IgG antibody reacts with the heavy chains of mouse IgG and the light chains common to most mouse immunoglobulins. No cross-reactivity has been detected against non-immunoglobulin serum proteins. This product exhibits minimal inherent cross-reaction with rat antigens. This antibody has been solid-phase absorbed to ensure minimal cross-reaction with rat, human, bovine, horse, and rabbit immunoglobulins, but may cross-react with other subclasses of mouse immunoglobulins. Poly $105 Anti-Rabbit IgG Poly4064 Reactivity: Rabbit 293E cells were stained by purified anti-tubulin-gamma (Poly6209) and labeled with BV421 anti-rabbit-igg (Poly4064 (blue)). Nuclei were stained with propidium iodide (red). Isotype: Donkey polyclonal Ig WB 1, IF, or FC (depending on format) Quality tested ELISA, IHC Reported in the literature This polyclonal donkey anti-rabbit IgG antibody reacts with the heavy chains of rabbit IgG and with the light chains common to most rabbit immunoglobulins. No cross-reactivity has been detected against non-immunoglobulin serum proteins. This antibody has been solid-phase absorbed to ensure minimal crossreaction with rat, mouse, human, bovine, horse, goat, chicken, guinea pig, Syrian hamster, and sheep immunoglobulins, but it may cross-react with other subclasses of rabbit immunoglobulins. Application References: 1. Lawson, B. R., et al J. Immunol. 178: Poly DyLight HRP μl DyLight Anti-Rat IgG1 Cy DyLight Clone: MRG1-58 Isotype: Mouse IgG Immunogen: Mixed rat Igs DyLight Brilliant Violet μl $185 Reactivity: Rat FC or ELISA (depending on format) Quality tested IHC, IF Reported in the literature The MRG1-58 monoclonal antibody reacts with rat immunoglobulin G1 (IgG1) in all tested rat strains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). It does not react with other isotypes. The MRG1-58 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone MRG Anti-Rat IgG2a Clone: MRG2a-83 Isotype: Mouse IgG Immunogen: Mixed rat Igs Reactivity: Rat FC 1 or ELISA (depending on format) Quality tested The MRG2a-83 monoclonal antibody reacts with rat immunoglobulin G2a (IgG2a) in all tested rat strains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). It does not react with other isotypes. The MRG2a-83 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Application References: 1. Inoue, S., et al Cancer Research 66: Clone MRG2a Anti-Rat IgG2b Clone: MRG2b-85 Isotype: Mouse IgG1, κ Immunogen: Mixed rat Igs Reactivity: Rat FC or ELISA (depending on format) Quality tested The MRG2b-85 monoclonal antibody reacts with rat immunoglobulin G2b (IgG2b) in all tested rat strains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). The MRG2b-85 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone MRG2b Anti-Rat IgG2c Clone: MRG2c-67 Isotype: Mouse IgG Immunogen: Mixed rat Igs Reactivity: Rat 558 Toll Free BIOLEGEND ( )

9 FC Reported in the literature The MRG2c-67 monoclonal antibody reacts with rat immunoglobulin G2c (IgG2c) in all tested rat strains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). It does not react with other isotypes. The MRG2c-67 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Application References: 1. Lee, J.H., et al J. Immunol. 178: 301. Clone MRG2c Anti-Rat IgM Clone: MRM-47 Isotype: Mouse IgG Immunogen: Mixed rat Igs ELISA Reported in the literature MRM-47 RT1D Alexa Fluor 647 LOU rat splenocytes stained with purified MRM-47 conjugated with and MHC class II antigen RT1D (14-4-4S) Alexa Fluor 647 The MRM-47 monoclonal antibody reacts with rat immunoglobulin M (IgM) in all tested rat strains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). It does not react with other isotypes. The MRM-47 antibody can be used as a primary or secondary reagent for ELISA or immunofluorescent analysis. Clone MRM $ $165 Anti-Rat Ig light chain κ Clone: MRK-81 Isotype: Mouse IgG Immunogen: Mixed rat Igs Reactivity: Rat FC Reported in the literature The MRK-81 monoclonal antibody reacts with rat immunoglobulin light chain kappa in all tested rat srains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). It does not react with lambda chain. The MRK-81 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone MRK Anti-Rat Ig light chain λ Clone: MRL-61 Isotype: Mouse IgG Immunogen: Mixed rat Igs Reactivity: Rat ELISA or FC (depending on format) Quality tested The MRL-61 monoclonal antibody reacts with rat immunoglobulin light chain lambda in all tested rat strains (Lou, Lou/Ws1/M, Lewis, Wistar, DA, Sprague-Dawley). It does not react with kappa chain. The MRL-61 monoclonal antibody may be used as primary or secondary reagent for ELISA or immunofluorescent analysis. Clone MRL Anti-Rat IgG Poly4054 Isotype: Goat polyclonal Ig Reactivity: Rat FC or ELISA (depending on format) Quality tested WB, IHC, IF (Cy3) Reported in the literature 293E cells were stained with purified anti-histone H3- Phospho (Ser28) antibody (clone HTA28) and labeled with BV421 anti-rat IgG (Poly4054) (blue). Cytosol is indicated by Alexa Fluor 488 anti-tubulinalpha (clone 10D8)(green) and nuclei are stained by propidium iodide. HTA28 only stains mitotic cells (violet). This polyclonal goat anti-rat IgG antibody reacts with the heavy chains of rat IgG and with the light chains common to most rat immunoglobulins. No crossreactivity has been detected against non-immunoglobulin serum proteins. This antibody has been solid-phase absorbed to ensure minimal cross-reaction with mouse, human, bovine, horse, and rabbit immunoglobulins, but it may cross-react with other subclasses of rat immunoglobulins. Poly $200 DyLight $110 HRP μl DyLight Cy AKP μl $180 DyLight Brilliant Violet μl $185 /Cy DyLight biolegend.com cs@biolegend.com 559

10 Avidin FC, IC Avidin binds to biotin with high affinity. Avidin-fluorescein isothiocyanate () is useful for detecting biotinylated antibodies. The excitation of by 488 nm laser light induces a light emission maximum of 520 nm. Avidin- is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. Avidin $ HRP Avidin ELISPOT, WB, IHC Reported in the literature Avidin binds to biotin with high affinity. Avidin-horseradish peroxidase (HRP) is useful for detecting biotinylated antibodies, e.g., in ELISA or Western blotting. For ELISA applications, HRP will act on soluble substrates, such as ABTS or TMB, to yield a colorimetric reaction. For Western blotting or ELISPOT applications, HRP will act on precipitating substrates, such as 4CN, to yield a colorimetric reaction. Avidin-HRP is useful as a second step reagent for indirect enzymatic labeling, in conjunction with biotinylated primary antibodies. Av-HRP is recommended for ELISA, ELISPOT, IHC, and Western blotting, when used with the relevant substrate system. Avidin HRP ml $110 AKP ELISPOT, WB, IHC Reported in the literature binds to biotin with high affinity. -AKP is useful for detecting biotinylated antibodies, e.g., in ELISA, ELISPOT, IHC or Western blotting. For ELISA applications, AKP will act on soluble substrates, such as pnpp, to yield a colorimetric reaction. For ELISPOT or Western blotting applications, AKP will act on precipitating substrates, such as NBT/BCIP, to yield a colorimetric reaction. -AKP is useful as a second step reagent for indirect enzymatic labelling, in conjunction with biotinylated primary antibodies. SAv-AKP is recommended for ELISA, ELISPOT, IHC, and Western blotting, when used with the relevant substrate system. AKP ml $110 FC, IC binds to biotin with high affinity. - is useful for detecting biotinylated antibodies. The excitation of by nm laser light induces a fluorescence emission maximum of 660 nm. -Allophycocyanin () is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. Application References: 1. Kamimura, D., et al J. Immunol. 177: Andoniou, C. E., et al Nature Immunology 6: /Cy7 FC, IC binds to biotin with high affinity. -Allophycocyanin/Cy7 (/Cy7) is useful for detecting biotinylated antibodies. The excitation of /Cy7 by nm laser light induces a fluorescence emission maximum of 780 nm. -/Cy7 is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. Application References: 1. Maresz, K., et al J. Neurochemistry 95: Ponomarev, E. D., et al J. Immunol. 176: Ponomarev, E. D., et al J. Immunol. 178: Mann, M. K., et al J. Immunol. 178: /Cy Brilliant Violet 421 Human peripheral blood lymphocytes were stained with biotinylated CD3 (filled histogram) or mouse IgG1 isotype control (open histogram), followed with SAV-Brilliant Violet 421. IF Validated binds to biotin with high affinity. The excitation of Brilliant Violet 421 by 405 nm laser light induces a fluorescence maximum emission of 421 nm. -Brilliant Violet 421 is useful as a second step reagent for indirect immunofluorescent staining when used in conjunction with biotinylated primary antibodies. 560 Toll Free BIOLEGEND ( )

11 Brilliant Violet μl $495 DyLight 549 IF Cy3 IF Quality tested binds to biotin with high affinity. -Cy3 is useful for detecting biotinylated antibodies. The excitation of Cy3 by 550 nm laser light induces a light emission maximum of 570 nm. Cy Cy5 FC, IC binds to biotin with high affinity. -Cy5 is useful for detecting biotinylated antibodies. The excitation of Cy5 by nm laser light induces a fluorescence emission maximum of 670 nm. -Cy5 is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. Application References: 1. Hsu, H., et al J. Immunol. 178: Cy DyLight 488 IF Reported in the literature binds to biotin with high affinity. The excitation of DyLight 488 by 493 nm laser light induces a fluorescence emission maximum of 518 nm. -DyLight 488 is useful as a second step reagent for indirect immunofluorescent staining when used in conjunction with biotinylated primary antibodies. Application References: 1. Ramos, TN., et al J. Immunol. 186:6657. DyLight E cell was stained with biotinylated anti-tubulin-alpha (clone 10D8) and labeled with -DyLight 549 (red). The nucleus of the cell was stained with DAPI (blue). DyLight DyLight 594 binds to biotin with high affinity. The excitation of DyLight 549 by 562 nm laser light induces a fluorescence emission maximum of 576 nm. -DyLight 549 is useful as a second step reagent for indirect immunofluorescent staining when used in conjunction with biotinylated primary antibodies. IF binds to biotin with high affinity. The excitation of DyLight 594 by 593 nm laser light induces a fluorescence emission maximum of 616 nm. -DyLight 594 is useful as a second step reagent for indirect immunofluorescent staining when used in conjunction with biotinylated primary antibodies. DyLight DyLight 649 IF Reported in the literature binds to biotin with high affinity. The excitation of DyLight 649 by 654 nm laser light induces a fluorescence emission maximum of 670 nm. -DyLight 649 is useful as a second step reagent for indirect immunofluorescent staining when used in conjunction with biotinylated primary antibodies. DyLight FC, IC binds to biotin with high affinity. -fluorescein isothiocyanate () is useful for detecting Continues biolegend.com cs@biolegend.com 561

12 , continued biotinylated antibodies. The excitation of by 488 nm laser light induces a light emission maximum of 520 nm. - is useful as a second step reagent for indirect immunofluorescent staining, in conjunction with biotinylated primary antibodies $ HRP ELISPOT, WB, IHC Reported in the literature binds to biotin with high affinity. -HRP is useful for detecting biotinylated antibodies, e.g., in ELISA, ELISPOT, IHC or Western blotting. For ELISA applications, HRP will act on soluble substrates, such as ABTS or TMB, to yield a colorimetric reaction. For ELISPOT or Western blotting applications, HRP will act on precipitating substrates, such as 4CN, to yield a colorimetric reaction. -HRP is useful as a second step reagent for indirect enzymatic labelling, in conjunction with biotinylated primary antibodies. SAv-HRP is recommended for ELISA, ELISPOT, IHC, and Western blotting, when used with the relevant substrate system. Application References: 1. Li, K-J., et al J. Leukoc. Biol. doi: / jlb Weldon, S., et al J. Immunol. 183: Basnayake, K., et al J Am Soc Naphrol. 21: Marder, W., et al Ann Rheum Dis. HRP ml $110 C57BL/6 splenocytes stained with 145-2C11 biotin, followed by SAv- FC, IC binds to biotin with high affinity. -Phycoerythrin () is useful for detecting biotinylated antibodies. The excitation of by 488 nm laser light induces a light emission maximum of 575 nm. -Phycoerythrin () is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. Application References: 1. Chappaz, S., et al Blood doi: / blood $ $160 /Cy5 C57BL/6 splenocytes stained with 145-2C11 biotin, followed by SAv-/Cy5 FC, IC binds to biotin with high affinity. -/Cy5 is useful for detecting biotinylated antibodies. The excitation of /Cy5 by 488 nm laser light induces a light emission maximum of 670 nm. -Phycoerythrin/Cy5 (/Cy5) is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. /Cy /Cy7 FC, IC binds to biotin with high affinity. -Phycoerythrin/Cy7 (/Cy7) is useful for detecting biotinylated antibodies. The excitation of /Cy7 by 488 nm laser light induces a light emission maximum of 775 nm. -/Cy7 is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. Application References: 1. Kamimura, D., et al J. Immunol. 177: Chappaz, S., et al Blood doi: /blood Anderson, S. M., et al J. Exp. Med. doi: /jem /Cy PerCP FC, IC binds to biotin with high affinity. -PerCP is useful for detecting biotinylated antibodies. The excitation of PerCP by 488 nm laser light induces a light emission maximum of 675 nm. -PerCP is useful as a second step reagent for indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. PerCP Toll Free BIOLEGEND ( )

13 PerCP/Cy5.5 Anti- FC, IC binds to biotin with high affinity. -PerCP/ Cy5.5 is useful for detecting biotinylated antibodies. The excitation of PerCP/Cy5.5 by 488 nm laser light induces a light emission maximum of 690 nm. C57BL/6 splenocytes stained -PerCP/Cy5.5 is useful as a second step reagent for with biotinylated 145-2C11, followed by Sav-PerCP/Cy5.5 indirect immunofluorescent staining, when used in conjunction with biotinylated primary antibodies. PerCP/Cy Anti-Allophycocyanin Clone: 003 Isotype: Mouse IgG1, κ Immunogen: Allophycocyanin Reactivity: Allophycocyanin Negative Control RM4-5 RM4-5 C57BL/6 thymocytes stained with RM4-5 only (left panel), or with RM4-5 plus biotinylated 003 (right panel), then detected with Sav- The 003 antibody reacts with both allophycocyanin () alone and conjugated with antibody or streptavidin, as well as tandem dyes such as /Cy5.5 and /Cy7. When 003 antibody is used in a three-step staining procedure or separation of -antibody labeled cells, it neither quenches fluorescence nor reacts with tested mouse and human cells. The 003 antibody can be used for separation of -antibody labeled cells or as a secondary step to amplify conjugated primary antibody signals (i.e., biotinylated 003 as secondary step, followed by -). Clone $ ylated 003+SAv- Clone: 1D4-C5 Isotype: Mouse IgG2a, κ Immunogen: ylated limulus hemocyanin IP, ELISA Reported in the literature C57BL/6 thymocytes stained with biotinylated CD4 (GK1.5), followed by purified 1D4-C5 and anti-mouse IgG (vitamin B7 or vitamin H) is essential for cell growth and the metabolization of fat and protein. It is also often used to tag a molecule of interest. The antibiotin antibody can be used as a second-step reagent for detecting biotinylated targets. Application References: 1. Uccellini, M.B., et al J. Immunol. 181:5875. Clone 1D4-C $85 LEAF $ $65 Anti-Fluorescein () Clone: FIT-22 FIT-22-anti-mouse IgG Negative Control RM4-5 C57BL/6 thymocytes stained with RM4-5 only (top) or RM4-5 plus purified FIT-22 (bottom), then detected with anti-mouse IgG Clone FIT RM Isotype: Mouse IgG1, κ Immunogen: Fluorescein Reactivity: Fluorescein The FIT-22 antibody reacts with both fluorescein () alone and conjugated with antibody or streptavidin. When FIT-22 antibody is used in a three-step staining procedure or separation of -antibody labeled cells, it does not quench fluorescence or react with tested mouse and human cells. The FIT-22 antibody can be used for separation of -antibody labeled cells or as a secondary step to amplify conjugated primary antibody signals (i.e. biotinylated FIT- 22 as secondary step, followed by -). LEAF : Low Endotoxin, Azide-Free biolegend.com cs@biolegend.com 563

14 Anti-KLH Anti-Phycoerythrin () Clone: MTK2059 Isotype: Mouse IgG1, κ Immunogen: Keyhole limpet hemocyanin (KLH) Reactivity: KLH The MTK2059 antibody is specific for KLH (keyhole limpet hemocyanin) alone or KLH conjugated protein. The MTK2059 antibody can be used for ELISA to detect KLH or KLH conjugates. Clone MTK Clone: MTK Isotype: Mouse IgG1, κ Immunogen: Keyhole limpet hemocyanin (KLH) Reactivity: KLH The MTK2113 antibody is specific for KLH (keyhole limpet hemocyanin) alone or KLH conjugated protein. The MTK2113 antibody can be used for ELISA to detect KLH or KLH conjugates. Clone MTK Clone: 001 ylated SAv- Negative Control Isotype: Mouse IgG1 Immunogen: Phycoerythrin Reactivity: Phycoerythrin The 001 antibody reacts with both phycoerythrin () alone and conjugated with antibody or streptavidin, as well RM4-5 as tandem dyes such as / Cy5, /Cy5.5 and /Cy7. When 001 antibody is used in a three-step staining procedure or separation of -antibody labeled cells, it does not quench fluorescence or react with tested mouse and human cells. RM4-5 The 001 antibody can be used C57BL/6 thymocytes stained with RM4-5 only (top) or for separation of -antibody labeled cells or as a secondary step RM4-5 plus biotinylated 001 (bottom), then to amplify conjugated primary antibody signals (i.e., bio- detected with Sav- tinylated 001 as secondary step, followed by -). Application References: 1. Taylor, R. T., et al J. Immunol. 178: Clone $ Toll Free BIOLEGEND ( )