Product Datasheet. c-myc Antibody (9E10) NB Unit Size: 0.1 ml

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1 Product Datasheet c-myc Antibody (9E10) NB Unit Size: 0.1 ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 3 Publications: 26 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 2/20/2019 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB c-myc Antibody (9E10) Product Information Unit Size Concentration Storage Clonality Clone Preservative Isotype Purity Buffer 0.1 ml 1 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Monoclonal 9E % Sodium Azide IgG1 Kappa Protein G purified PBS Page 1 of 8 v.20.1 Updated 2/20/2019 Product Description Host Mouse Gene ID 4609 Gene Symbol Species Reactivity Notes Specificity/Sensitivity MYC Human, Mouse, Drosophila Human, mouse and Drosophila. Specific for the c-myc protein in random coil configuration, not as a helix. 9E10 does not react with V-myc. Immunogen A synthetic peptide corresponding to amino acids (AEEQKLISEEDLLRKRREQLKHKLEQLRNSCA) of human c-myc. [UniProt# P01106] Product Application Details Applications Recommended Dilutions Western Blot, Simple Western, Chromatin Immunoprecipitation, ELISA, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry- Paraffin, Immunoprecipitation Western Blot ug/ml, Simple Western 1:200, Chromatin Immunoprecipitation, Flow Cytometry 1:50-1:200, ELISA 1:100-1:2000, Immunohistochemistry 1:50-1:200, Immunocytochemistry/Immunofluorescence 1:50-1:200, Immunoprecipitation 1:10-1:500, Immunohistochemistry-Paraffin 1:50-1:200, Immunohistochemistry-Frozen 1:50-1:200, Flow (Intracellular) Application Notes This c-myc antibody clone 9E10 is useful for Flow Cytometry (PMID: ), Western Blot, ELISA, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry- Paraffin and Immunoprecipitation. In Simple Western only ul of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.

3 Images Western Blot: c-myc Antibody (9E10) [NB ] - Analysis of c-myc in Jurkat cell lysates using NB Page 2 of 8 v.20.1 Updated 2/20/2019 Immunohistochemistry-Paraffin: c-myc Antibody (9E10) [NB ] - c- Myc was detected in immersion fixed paraffin-embedded sections of human breast cancer using anti-human mouse monoclonal antibody (Catalog # NB , clone 9E10) at 1:50 dilution overnight at 4 C. Tissue was stained using the VisuCyte anti-mouse HRP polymer detection reagent (Catalog # VC001) with DAB chromogen (brown) and counterstained with hematoxylin (blue). Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand. Flow Cytometry: c-myc Antibody (9E10) [NB ] - Analysis of Alexa Fluor (R) 647 conjugate of NB An intracellular stain was performed on HeLa cells with c-myc antibody (9E10) NB AF647 (blue) and a matched isotype control NBP AF647 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to AF647 Immunocytochemistry/Immunofluorescence: c-myc Antibody (9E10) [NB ] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-c-myc (9E10) at 10 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

4 stain was performed on HeLa cells with c-myc Antibody [9E10] NB C (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650. Page 3 of 8 v.20.1 Updated 2/20/2019 Flow Cytometry: c-myc Antibody (9E10) [NB ] - Analysis of transgenic Eu-Myc+ mouse splenocytes using c-myc PE conjugated antibody. Image from verified customer review. Flow (Intracellular): c-myc Antibody (9E10) [NB ] - An intracellular stain was performed on U-937 cells with c-myc Antibody (9E10) NB F (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to FITC. Flow (Intracellular): c-myc Antibody (9E10) [NB ] - An intracellular stain was performed on U-937 cells with c-myc Antibody (9E10) NB PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin.

5 stain was performed on U-937 cells with c-myc Antibody (9E10) NB AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488. Page 4 of 8 v.20.1 Updated 2/20/2019 stain was performed on HeLa cells with c-myc Antibody (9E10) NB and a matched isotype control NBP Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/ml for 30 minutes at room temperature, followed by Mouse F(ab)2 IgG (H+L) PE-conjugated secondary antibody (F0102B, R&D Systems). Flow Cytometry: c-myc Antibody (9E10) [NB ] - Gray: WT B cell control Blue: EuMyc un-transformed B cells Red: EuMyc lymphoma This Image was submitted by customer review. Image from the FITC version of this antibody. stain was performed on A431 cells with c-myc Antibody (9E10) NB AF700 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 700.

6 Flow Cytometry: c-myc Antibody (9E10) [NB ] - Gray: WT B cell control. Blue: EuMyc un-transformed B cells. Red: EuMyc lymphoma. This image was submitted via customer review. Image from the FITC version of this antibody. Page 5 of 8 v.20.1 Updated 2/20/2019 stain was performed on Jurkat cells with c-myc Antibody (9E10) NB R (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/ml for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550. Simple Western: c-myc Antibody (9E10) [NB ] - Simple Western lane view shows a specific band for c-myc in 0.5 mg/ml of Jurkat lysate. This experiment as performed under reducing conditions using the kda separation system.

7 Publications Morioka S, Perry JSA, Raymond MH et al. Efferocytosis induces a novel SLC program to promote glucose uptake and lactate release. Nature. Nov :00AM [PMID: ] (ICC/IF, Mouse) Rodriguez-Muela N, Parkhitko A, Grass T et al. Blocking p62/sqstm1-dependent SMN degradation ameliorates Spinal Muscular Atrophy disease phenotypes. J. Clin. Invest. Apr :00AM [PMID: ] (Mouse) van der Kant R, Jonker CT, Wijdeven RH et al. Characterization of the Mammalian CORVET and HOPS Complexes and Their Modular Restructuring for Endosome Specificity J Biol Chem 2015 Dec 18 [PMID: ] (Human) Details: This reference used the HRP version of NB Mahadevan S, Sathappan V, Utama B et al. Maternally expressed NLRP2 links the subcortical maternal complex (SCMC) to fertility, embryogenesis and epigenetic reprogramming. Sci Rep. Mar :00AM [PMID: ] (ICC/IF, Mouse) Hsieh YT, Chou MM, Chen HC, Tseng JJ. IMP1 promotes choriocarcinoma cell migration and invasion through the novel effectors RSK2 and PPME1. Gynecol Oncol Oct [PMID: ] Jang KJ, Mano H, Aoki K et al. Mitochondrial function provides instructive signals for activation-induced B-cell fates Nat Commun. Nat Commun 2015 Apr 10 [PMID: ] (WB, Mouse) Details: Using the DyLight 650 conjugated version of NB , catalog number NB C. da Costa JP, Carvalhais V, Amado F et al. Anti-tumoral activity of human salivary peptides Peptides Jul 23 [PMID: ] (WB, Human) Details: c-myc antibody used for Western blot on human cell lines from MG63, HT29, T24 and T-47 cells that were treated with different concentrations of GG peptide (Fig 4). Iampietro M, Morissette G, Gravel A, Flamand L. Inhibition of Interleukin-2 Gene Expression by Human Herpesvirus 6B U54 Tegument Protein. J Virol Aug 13 [PMID: ] (WB, Human) Pekkurnaz G, Trinidad JC, Wang X et al. Glucose Regulates Mitochondrial Motility via Milton Modification by O- GlcNAc Transferase. Cell 2014 Jul 03 [PMID: ] (ICC/IF) Details: COS7 cells transfected with empty vector or myc-hmilton1 (Figure S3). Barnett TC, Liebl D, Seymour LM et al. The Globally Disseminated M1T1 Clone of Group A Streptococcus Evades Autophagy for Intracellular Replication. Cell Host Microbe 2013 Dec 11 [PMID: ] (ICC/IF) Hubbi ME, Hu H, Kshitiz NF et al. Sirtuin-7 Inhibits the Activity of Hypoxia-Inducible Factors J Biol Chem 2013 June 9 [PMID: ] Megill A, Lee T, Dibattista AM et al. A Tetra(Ethylene Glycol) Derivative of Benzothiazole Aniline Enhances Ras- Mediated Spinogenesis. J Neurosci 2013 May 29 [PMID: ] (ICC/IF) More publications at Page 6 of 8 v.20.1 Updated 2/20/2019

8 Procedures Page 7 of 8 v.20.1 Updated 2/20/2019 Protocol specific for c-myc Antibody (NB ) Western Blot Protocol 1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 25 ug of total protein per lane. 2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus. 3. Rinse membrane with dh2o and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil. 4. Rinse the blot in TBS for approximately 5 minutes. 5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT. 6. Rinse the membrane in dh2o and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each. 7. Dilute the mouse anti-c-myc primary antibody (NB ) in blocking buffer and incubate 1 hour at room temperature. 8. Rinse the membrane in dh2o and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each. 9. Apply the diluted mouset-igg HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature. 10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background). 11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL). Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of %, provided it does not interfere with antibody-antigen binding.

9 Novus Biologicals USA E. Briarwood Avenue Centennial, CO USA Phone: Toll Free: Fax: Novus Biologicals Canada 461 North Service Road West, Unit B37 Oakville, ON L6M 2V5 Canada Phone: Toll Free: Fax: Novus Biologicals Europe 19 Barton Lane Abingdon Science Park Abingdon, OX14 3NB, United Kingdom Phone: (44) (0) Free Phone: Fax: (44) (0) General Contact Information Technical Support: Orders: General: Products Related to NB NB800-PC2 HAF007 NB720-B NBP mg Jurkat Whole Cell Lysate Goat anti-mouse, IgG Secondary Antibody [HRP (Horseradish Peroxidase)] Rabbit anti-mouse IgG (H+L) Secondary Antibody [Biotin] Mouse IgG1 Kappa Light Chain Isotype Control (P ) Limitations This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt. For more information on our 100% guarantee, please visit Earn gift cards/discounts by submitting a review: Earn gift cards/discounts by submitting a publication using this product: