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BM4008S Macrophage F4/80 antigen antibody

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0.25 mg / €350.00

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Rat anti Mouse Macrophage F4/80 antigen Cl:A3-1

BM4008S

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Product Description for Macrophage F4/80 antigen

Rat anti Mouse Macrophage F4/80 antigen Cl:A3-1.
Presentation: Purified
Product is tested for Immunoelectron Microscopy, Frozen Sections, Immunoprecipitation, Radioimmunoassay, Western blot / Immunoblot, Flow Cytometry, Paraffin Sections.

Properties for Macrophage F4/80 antigen

Product Category Primary Antibodies
Target Category
Quantity 0.25 mg
Synonyms Cell surface glycoprotein EMR1, EMR1 hormone receptor, Emr1, Gpf480
Presentation Purified
Reactivity Ms
Applications C, EM, F, IP, P, R, WB
Clonality Monoclonal
Clone Cl:A3-1
Host Rat
Isotype IgG2b
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen
Swiss Prot Num:
Q61549
Immunogen:
Thioglycollate stimulated peritoneal macrophages from C57/BL mice.
Spleen cells from immunised HOB2 rats were fused with cells of the mouse NS1 myeloma cell line.
GeneID:
13733
Isotype control SM19P
Application RIA.
Western Blot.
Immunoprecipitation.
Immunofluorescence. 
Immunoelectron Microscopy. 

Flow Cytometry: Use 10 µl of 1/50-1/100 diluted antibody to label 106 cells in 100 µl.
Immunohistochemistry on Frozen and Paraffin Embedded and Resin Sections. This product requires pre-treatment of paraffin sections (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).
Background F4/80 antigen is a 160 kD glycoprotein expressed by most murine macrophages.
Expression of F4/80 is heterogeneous and is reported to vary during macrophage maturation and activation. The F4/80 antigen is expressed on a wide range of mature tissue macrophages including Kupffer cells, Langerhans, microglia, macrophages located in the gut lamina propria, peritoneal cavity, lung, thymus, bone marrow stroma and macrophages in the red pulp of the spleen. F4/80 expression has also been reported on a subpopulation of dendritic cells but is absent from macrophages located in T cell areas of the spleen and lymphnode. The ligands and biological functions of the F4/80 antigen have not yet been determined but recent studies suggest a role for F4/80 in the generation of efferent CD8+ve regulatory T cells.
Concentration 1.0 mg/ml
Protocols 1. Enzyme pre-treatment using Proteinase K (Recommended for tissues fixed for 24 hours in neutral buffered formalin, NBF):
Reagents
A. TE buffer (50 mM Tris base, 1 mM EDTA, pH 8.0)
Tris Base, 6.10 g
EDTA, 0.37 g
Distilled water, 1000 ml
Mix to dissolve. Adjust pH to 8.0 using concentrated HCl (10 M HCl). Store at room temperature.

B. Proteinase K stock solution (20x, 400 μg/ml in TE buffer, pH 8.0)
Proteinase K, 4 mg
TE buffer, pH 8.0, (Reagent A) 10 ml
Mix well. Store in aliquots at -20°C.

C. Proteinase K working solution (1x, 20 μg/ml in TE buffer, pH 8.0)
Proteinase K stock solution (20x), (Reagent B) 1 ml
TE Buffer, pH 8.0, (Reagent A) 19 ml
Mix well. Discard working solution after use.

Method
1. Dewax paraffin sections and rehydrate using preferred procedure.
2. Cover sections completely with Proteinase K working solution and incubate for 3 minutes at RT.
3. Rinse sections with Phosphate Buffered Saline (PBS).
4. Proceed with serum blocking and preferred staining protocol.

2. Heat-mediated antigen retrieval using citrate buffer, pH 6.0 (Recommended for tissues fixed for 7 days or more in neutral buffered formalin, NBF):
Reagent
Citrate buffer (10 mM citric acid, pH 6.0)

Citric acid (anhydrous), 1.92 g
Distilled water, 1000 ml
Mix to dissolve. Adjust pH to 6.0 with 1 M NaOH (be sure to mix well). Store this solution at RT for 3 months, or at 4°C for longer usage.

Method
1. Dewax paraffin sections and rehydrate using preferred protocol.
2. Pre-heat sodium citrate buffer in a staining vessel to 95-100°C.
3. Immerse slides in the citrate buffer and incubate for 10 minutes at 95-100°C. Check the citrate buffer level, add more if necessary, and then incubate for a further 10 minutes at 95-100°C.
4. Allow sections to cool for 20 minutes.
5. Rinse sections with PBS.
6. Proceed with serum blocking and preferred staining protocol.
Product Citations
Purchased from Acris:
  1. Kakehashi H, Nishioku T, Tsukuba T, Kadowaki T, Nakamura S, Yamamoto K. Differential regulation of the nature and functions of dendritic cells and macrophages by cathepsin E. J Immunol. 2007 Nov 1;179(9):5728-37. PubMed PMID: 17947645.
  2. Kimura H, Miyashita H, Suzuki Y, Kobayashi M, Watanabe K, Sonoda H, et al. Distinctive localization and opposed roles of vasohibin-1 and vasohibin-2 in the regulation of angiogenesis. Blood. 2009 May 7;113(19):4810-8. doi: 10.1182/blood-2008-07-170316. Epub 2009 Feb 9. PubMed PMID: 19204325.
  3. Cuzić S, Bosnar M, Kramarić MD, Ferencić Z, Marković D, Glojnarić I, et al. Claudin-3 and Clara cell 10 kDa protein as early signals of cigarette smoke-induced epithelial injury along alveolar ducts. Toxicol Pathol. 2012 Dec;40(8):1169-87. doi: 10.1177/0192623312448937. Epub 2012 Jun 1. PubMed PMID: 22659244.
Originator or purchased from resellers:
  1. Nakao S, Zandi S, Faez S, Kohno R, Hafezi-Moghadam A. Discontinuous LYVE-1 expression in corneal limbal lymphatics: dual function as microvalves and immunological hot spots. FASEB J. 2012 Feb;26(2):808-17. doi: 10.1096/fj.11-183897. Epub 2011 Nov 16. PubMed PMID: 22090317. (Free PMC Article available, 7 images available)
  2. Anghelina M, Krishnan P, Moldovan L, Moldovan NI. Monocytes/macrophages cooperate with progenitor cells during neovascularization and tissue repair: conversion of cell columns into fibrovascular bundles. Am J Pathol. 2006 Feb;168(2):529-41. PubMed PMID: 16436667. (Free PMC Article available, 8 images available)
General Readings
  1. Austyn JM, Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805-15. PubMed PMID: 7308288.
  2. Hume DA, Perry VH, Gordon S. The mononuclear phagocyte system of the mouse defined by immunohistochemical localisation of antigen F4/80: macrophages associated with epithelia. Anat Rec. 1984 Nov;210(3):503-12. PubMed PMID: 6524692.
  3. Lee SH, Starkey PM, Gordon S. Quantitative analysis of total macrophage content in adult mouse tissues. Immunochemical studies with monoclonal antibody F4/80. J Exp Med. 1985 Mar 1;161(3):475-89. PubMed PMID: 3973536. (Free PMC Article available)
  4. Gordon S, Lawson L, Rabinowitz S, Crocker PR, Morris L, Perry VH. Antigen markers of macrophage differentiation in murine tissues. Curr Top Microbiol Immunol. 1992;181:1-37. PubMed PMID: 1424778.
  5. Warschkau H, Kiderlen AF. A monoclonal antibody directed against the murine macrophage surface molecule F4/80 modulates natural immune response to Listeria monocytogenes. J Immunol. 1999 Sep 15;163(6):3409-16. PubMed PMID: 10477612.
  6. Lin HH, Faunce DE, Stacey M, Terajewicz A, Nakamura T, Zhang-Hoover J, et al. The macrophage F4/80 receptor is required for the induction of antigen-specific efferent regulatory T cells in peripheral tolerance. J Exp Med. 2005 May 16;201(10):1615-25. Epub 2005 May 9. PubMed PMID: 15883173. (Free PMC Article available, 6 images available)
  7. Chan RJ, Leedy MB, Munugalavadla V, Voorhorst CS, Li Y, Yu M, et al. Human somatic PTPN11 mutations induce hematopoietic-cell hypersensitivity to granulocyte-macrophage colony-stimulating factor. Blood. 2005 May 1;105(9):3737-42. Epub 2005 Jan 11. PubMed PMID: 15644411. (Free PMC Article available, 5 images available)
  8. Pizza FX, Peterson JM, Baas JH, Koh TJ. Neutrophils contribute to muscle injury and impair its resolution after lengthening contractions in mice. J Physiol. 2005 Feb 1;562(Pt 3):899-913. Epub 2004 Nov 18. PubMed PMID: 15550464. (Free PMC Article available, 8 images available)
  9. Metwali A, Blum AM, Elliott DE, Setiawan T, Weinstock JV. Cutting edge: hemokinin has substance P-like function and expression in inflammation. J Immunol. 2004 Jun 1;172(11):6528-32. PubMed PMID: 15153465.
  10. Moore KJ, Andersson LP, Ingalls RR, Monks BG, Li R, Arnaout MA, et al. Divergent response to LPS and bacteria in CD14-deficient murine macrophages. J Immunol. 2000 Oct 15;165(8):4272-80. PubMed PMID: 11035061.
  11. Hemmi H, Idoyaga J, Suda K, Suda N, Kennedy K, Noda M, et al. A new triggering receptor expressed on myeloid cells (Trem) family member, Trem-like 4, binds to dead cells and is a DNAX activation protein 12-linked marker for subsets of mouse macrophages and dendritic cells. J Immunol. 2009 Feb 1;182(3):1278-86. PubMed PMID: 19155473. (Free PMC Article available, 7 images available)
  12. Seitz O, Schürmann C, Hermes N, Müller E, Pfeilschifter J, Frank S, et al. Wound healing in mice with high-fat diet- or ob gene-induced diabetes-obesity syndromes: a comparative study. Exp Diabetes Res. 2010;2010:476969. doi: 10.1155/2010/476969. Epub 2011 Jan 20. PubMed PMID: 21318183. (Free PMC Article available, 8 images available)
  13. Miao EA, Leaf IA, Treuting PM, Mao DP, Dors M, Sarkar A, et al. Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria. Nat Immunol. 2010 Dec;11(12):1136-42. doi: 10.1038/ni.1960. Epub 2010 Nov 7. PubMed PMID: 21057511. (Free PMC Article available, 7 images available)
  14. Wang X, Yang Z, Xue B, Shi H. Activation of the cholinergic antiinflammatory pathway ameliorates obesity-induced inflammation and insulin resistance. Endocrinology. 2011 Mar;152(3):836-46. doi: 10.1210/en.2010-0855. Epub 2011 Jan 14. PubMed PMID: 21239433. (Free PMC Article available, 6 images available)
  15. Cunningham O, Campion S, Perry VH, Murray C, Sidenius N, Docagne F, et al. Microglia and the urokinase plasminogen activator receptor/uPA system in innate brain inflammation. Glia. 2009 Dec;57(16):1802-14. doi: 10.1002/glia.20892. PubMed PMID: 19459212. (Free PMC Article available, 5 images available)
  16. Gornicka A, Fettig J, Eguchi A, Berk MP, Thapaliya S, Dixon LJ, et al. Adipocyte hypertrophy is associated with lysosomal permeability both in vivo and in vitro: role in adipose tissue inflammation. Am J Physiol Endocrinol Metab. 2012 Sep 1;303(5):E597-606. doi: 10.1152/ajpendo.00022.2012. Epub 2012 Jun 26. PubMed PMID: 22739104. (Free PMC Article available, 6 images available)
  17. Akbarshahi H, Menzel M, Posaric Bauden M, Rosendahl A, Andersson R. Enrichment of murine CD68+ CCR2+ and CD68+ CD206+ lung macrophages in acute pancreatitis-associated acute lung injury. PLoS One. 2012;7(10):e42654. doi: 10.1371/journal.pone.0042654. Epub 2012 Oct 22. PubMed PMID: 23110041. (Free PMC Article available, 7 images available)
  18. Banda NK, Hyatt S, Antonioli AH, White JT, Glogowska M, Takahashi K, et al. Role of C3a receptors, C5a receptors, and complement protein C6 deficiency in collagen antibody-induced arthritis in mice. J Immunol. 2012 Feb 1;188(3):1469-78. doi: 10.4049/jimmunol.1102310. Epub 2011 Dec 28. PubMed PMID: 22205026. (Free PMC Article available, 7 images available)
  19. Bonde AK, Tischler V, Kumar S, Soltermann A, Schwendener RA. Intratumoral macrophages contribute to epithelial-mesenchymal transition in solid tumors. BMC Cancer. 2012 Jan 24;12:35. doi: 10.1186/1471-2407-12-35. PubMed PMID: 22273460. (Free PMC Article available, 6 images available)
  20. Choi KM, Kashyap PC, Dutta N, Stoltz GJ, Ordog T, Shea Donohue T, et al. CD206-positive M2 macrophages that express heme oxygenase-1 protect against diabetic gastroparesis in mice. Gastroenterology. 2010 Jun;138(7):2399-409, 2409.e1. doi: 10.1053/j.gastro.2010.02.014. Epub 2010 Feb 20. PubMed PMID: 20178793. (Free PMC Article available, 7 images available)
  21. Tamaki S, Mano T, Sakata Y, Ohtani T, Takeda Y, Kamimura D, et al. Interleukin-16 promotes cardiac fibrosis and myocardial stiffening in heart failure with preserved ejection fraction. PLoS One. 2013 Jul 19;8(7):e68893. doi: 10.1371/journal.pone.0068893. Print 2013. PubMed PMID: 23894370. (Free PMC Article available, 7 images available)
  22. Kihira Y, Miyake M, Hirata M, Hoshina Y, Kato K, Shirakawa H, et al. Deletion of hypoxia-inducible factor-1α in adipocytes enhances glucagon-like peptide-1 secretion and reduces adipose tissue inflammation. PLoS One. 2014 Apr 4;9(4):e93856. doi: 10.1371/journal.pone.0093856. eCollection 2014. PubMed PMID: 24705496. (Free PMC Article available, 5 images available)
  23. Chinzei N, Hayashi S, Ueha T, Fujishiro T, Kanzaki N, Hashimoto S, et al. P21 deficiency delays regeneration of skeletal muscular tissue. PLoS One. 2015 May 5;10(5):e0125765. doi: 10.1371/journal.pone.0125765. eCollection 2015. PubMed PMID: 25942471. (Free PMC Article available)
  24. Sumiyoshi, M. et al. (2015) Antitumor and antimetastatic actions of xanthoangelol and 4-hydroxyderricin isolated from Angelica keiskei roots through the inhibited activation and differentiation of M2 macrophages. Phytomedicine. 22 (7-8): 759-67.
Storage Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Format
Purification:
Affinity Chromatography on Protein G
Buffer System:
PBS, pH 7.4
Preservatives:
0.09% Sodium Azide
State:
Liquid purified IgG fraction
Purified
Species Reactivity
Species reactivity (tested):
Mouse.
Specificity
Specificity:
This antibody recognizes the F4/80 antigen, a member of the EGF-TM7 family of proteins which shares 68% overall amino acid identity with Human EMR1.
Clone CI:A31 has been reported to modulate cytokine levels released in response to Listeria monocytogenes (Ref.5). We recommend the use of BM4008LE for this purpose.
Gene ID 13733
FocusOn and Reviews
FocusOns:

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