Product Datasheet. F4/80 Antibody (CI-A3-1) NB Unit Size: mg

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1 Product Datasheet F4/80 Antibody (CI-A3-1) NB Unit Size: mg Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 1 Publications: 30 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 8/22/2018 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB F4/80 Antibody (CI-A3-1) Product Information Unit Size Concentration Storage Clonality Clone Preservative Isotype Purity Buffer Target Molecular Weight mg 1.0 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Monoclonal CI-A % Sodium Azide IgG2b Protein G purified Phosphate buffered saline 160 kda Page 1 of 7 v.20.1 Updated 8/22/2018 Product Description Host Rat Gene ID Gene Symbol Species ADGRE1 Human, Mouse Reactivity Notes Human reactivity reported in scientific literature (PMID: ). Marker Specificity/Sensitivity Immunogen Macrophage Marker This recognizes mouse F4/80, a 160kD glycoprotein expressed by murine macrophages. Thioglycollate stimulated peritoneal macrophages from C57/BL mice. Product Application Details Applications Recommended Dilutions Application Notes Western Blot, Electron Microscopy, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Radioimmunoassay Western Blot 1:100-1:2000, Flow Cytometry 1:50-1:100, Immunohistochemistry 1:10-1:500, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunoprecipitation 1:10-1:500RadioImmunoassay 1:100-1:2000, Immunohistochemistry-Paraffin 1:10-1:500, Immunohistochemistry-Frozen 1:10-1:500, Radioimmunoassay 1:100-1:2000, Electron Microscopy 1:10-1:500 This product requires pretreatment of paraffin sections prior to staining. Proteinase K is recommended for tissues fixed for less than 24 hours. Citrate buffer ph 6.0 is recommended for tissues fixed for more than 24 hours.

3 Images Immunocytochemistry/Immunofluorescence: F4/80 Antibody (CI-A3-1) [NB ] - Lymph node stained for macrophages, red and B cells CD79b, green; nuclei are stained with DAPI, blue. Page 2 of 7 v.20.1 Updated 8/22/2018 Immunocytochemistry/Immunofluorescence: F4/80 Antibody (CI-A3-1) [NB ] - Staining of mouse small intestine. Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) [NB ] - Mouse spleen stained with A: Rat anti Mouse. B: CI-A3-1 clone preincubated with 2 molar excess of human anti Idiotypic. Immunohistochemistry-Frozen: F4/80 Antibody (CI-A3-1) [NB ] - Frozen mouse spleen with F4/80 anti-mouse antibody.

4 Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB ] - Staining of J774 cells with F4/80 antibody. Page 3 of 7 v.20.1 Updated 8/22/2018 Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB ] - Staining of mouse peritoneal macrophages with F4/80 antibody. Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB ] - Staining of mouse peritoneal macrophages with clone CI-A3-1 F4/80 antibody. Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB ] - A surface stain was performed on RAW cells with F4/80 antibody (CI-A3-1) NB AF488 (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 10 ug/ml for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

5 Flow Cytometry: F4/80 Antibody (CI-A3-1) [NB ] - Staining of mouse peritoneal macrophages with F4/80 antibody. Page 4 of 7 v.20.1 Updated 8/22/2018

6 Publications Jin Yiping, Arita Makoto, Zhang Qiang et al. Anti-angiogenesis effect of the novel anti-inflammatory and pro-resolving lipid mediators. Investigative Ophthalmology & Visual Science 2009 [PMID: ] (IHC, Mouse) Lietman C, Wu B, Lechner S, Shinar A. Inhibition of Wnt/b-catenin signaling ameliorates osteoarthritis in a murine model of experimental osteoarthritis. JCI Insight Feb 08 [PMID: ] (IHC-P, Mouse) Iacobini C, Fantauzzi CB, Bedini R, Pecci R. Galectin-3 is essential for proper bone cell differentiation and activity, bone remodeling and biomechanical competence in mice. Metab. Clin. Exp Feb 09 [PMID: ] (IHC, Mouse) Zhang H, Lu H, Xiang L et al. HIF-1 regulates CD47 expression in breast cancer cells to promote evasion of phagocytosis and maintenance of cancer stem cells. Proc Natl Acad Sci Nov 10 [PMID: ] (FLOW, Human) Details: This citation used the Allophycocyanin form of this antibody. Moon H, Donahue LR, Choi E et al. Melanocyte Stem Cell Activation and Translocation Initiate Cutaneous Melanoma in Response to UV Exposure. Cell Stem Cell Oct 04 [PMID: ] (WB) Potula HH, Richer L, Werts C, Gomes-Solecki M. Pre-treatment with Lactobacillus plantarum prevents severe pathogenesis in mice infected with Leptospira interrogans and may be associated with recruitment of myeloid cells PLoS Negl Trop Dis 2017 Aug 25 [PMID: ] (IHC-P, Mouse) Yuan H, Jiang W, von Roemeling CA et al. Multivalent bi-specific nanobioconjugate engager for targeted cancer immunotherapy. Nat Nanotechnol May :00AM [PMID: ] (IHC-P) Chung KJ, Chatzigeorgiou A, Economopoulou M et al. A self-sustained loop of inflammation-driven inhibition of beige adipogenesis in obesity. Nat. Immunol. Apr :00AM [PMID: ] Wang W, Xu MJ, Cai Y et al. Inflammation is independent of steatosis in a murine model of steatohepatitis.. Hepatology. Feb :00AM [PMID: ] (IHC-P, Mouse) Demonbreun AR, Allen MV, Warner JL et al. Enhanced Muscular Dystrophy from Loss of Dysferlin Is Accompanied by Impaired Annexin A6 Translocation after Sarcolemmal Disruption. Am. J. Pathol Apr 09 [PMID: ] (IHC-Fr, ICC/IF, Mouse) Details: This publication used the AF488 conjugated form of this antibody (NB AF488) Merches K, Khairnar V, Knuschke T et al. Virus-Induced Type I Interferon Deteriorates Control of Systemic Pseudomonas Aeruginosa Infection Cell Physiol Biochem (IHC-Fr, Mouse) Details: This publication used the DyLight 488 conjugated form of this antibody (Cat# NB G). Merches K, Khairnar V, Knuschke T et al. Virus-Induced Type I Interferon Deteriorates Control of Systemic Pseudomonas Aeruginosa Infection Cell Physiol Biochem 2015 Aug 17 [PMID: ] (IHC-Fr, Mouse) Details: This citation used the DyLight 488 version of this antibody. More publications at Page 5 of 7 v.20.1 Updated 8/22/2018

7 Procedures Western Blot protocol for F4/80 Antibody (NB ) Western Blot Protocol Page 6 of 7 v.20.1 Updated 8/22/ Perform SDS-PAGE on samples to be analyzed, loading 30 ug of total protein per lane. 2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus. 3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate. 4. Rinse the blot. 5. Block the membrane using standard blocking buffer for at least 1 hour. 6. Wash the membrane in wash buffer three times for 10 minutes each. 7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature. 8. Wash the membrane in wash buffer three times for 10 minutes each. 9. Apply the diluted 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 three 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. **Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of %. Immunocytochemistry/Immunofluorescence Protocol for F4/80 Antibody (NB ) Immunocytochemistry Protocol Culture cells to appropriate density in 35 mm culture dishes or 6-well plates. 1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes. 2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes. 3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes. 4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature. 5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature. 6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes. 7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature. 8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes. 9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing. *The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

8 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 HAF005 Goat anti-rat IgG Secondary Antibody [HRP (Horseradish Peroxidase)] NB7115 Goat anti-rat IgG (H+L) Secondary Antibody [HRP] DDXCR03 Rat IgG2b Isotype Control NB AF488 F4/80 Antibody (CI-A3-1) [Alexa Fluor 488] 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: