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Safety Data Sheet (SDS) Brilliant Violet 421 Anti-mouse F4/80 Antibody     Product Data Sheet (PDF)    
Brilliant Violet 421™ Anti-mouse F4/80 Antibody
1215655 125 µl $180.00       
1215685 50 µg $235.00       
1215660 500 µl $365.00       
Clone: BM8
Isotype: Rat IgG2a, κ
Reactivity: Mouse
Immunogen: Murine macrophages
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation: The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 421™ under optimal conditions. The solution is free of unconjugated Brilliant Violet 421™ and unconjugated antibody.
Concentration: µg sizes: 0.2 mg/ml
µl sizes: lot-specific (please contact technical support for concentration and total µg amount)
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

FC - Quality tested
IHC-F - Validated

Application Notes:

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections1,2 and formalin-fixed paraffin-embedded sections6,7, and Western blotting.

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining using the µg size, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. For immunofluorescent staining using µl sizes, the suggested use of this reagent is ≤5 µl per million cells or 5 µl per 100 µl of whole blood. For immunohistochemical staining on frozen tissue sections, the suggested use of this reagent is 2.0 µg/ml. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.

Application References:

1. Schaller E, et al. 2002. Mol. Cell. Biol. 22:8035. (IHC)
2. Stevceva L, et al. 2001. BMC Clin Pathol. 1:3. (IHC)
3. Kobayashi M, et al.2008. J. Leukoc. Biol. 83:1354. PubMed
4. Poeckel D, et al. 2009. J. Biol Chem. 284:21077. PubMed
5. Glass AM, et al. 2013. J. Immunol. 190:4830. PubMed
6. Koehm S, et al. 2007. J. Allergy Clin. Immunol. 120:570. (IHC)
7. Rankin AL, et al. 2010. J. Immunol. 184:1526. (IHC)
8. Sasi SP, et al. 2014. J Biol Chem. 289:14178. PubMed
9. Thakus VS, et al. 2014. Toxicol Lett. 230:322. PubMed
10. Karsten CM, et al. 2015. J Immunol. 194:1841. PubMed

Thioglycolate-elicited Balb/c mouse peritoneal macrophages

Thioglycolate-elicited Balb/c mouse peritoneal macrophages were stained with F4/80 (clone BM8) Brilliant Violet 421™ (filled histogram) or rat IgG2a, κ Brilliant Violet 421™ isotype control (open histogram).

C57BL/6 mouse frozen spleen section

C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for 30 minutes at room temperature. Then the section was stained with 2 µg/ml of anti-mouse F4/80 (clone BM8) Brilliant Violet 421™ (Blue) and anti-mouse CD8a (clone 53-6.7) Alexa Fluor® 647 (red) overnight at 4°C. The image was captured with a 10X objective.



Description:

F4/80 is a 160 kD glycoprotein. It is characterized as a member of the epidermal growth factor (EGF)-transmembrane 7 (TM7) family. F4/80, also known as EMR1 or Ly71, has been widely used as a murine macrophage marker, which is expressed on the majority of tissue macrophages including peritoneal macrophages, macrophages in lung, gut, thymus and red pulp of spleen (but not on the macrophages located in T cell areas of the spleen, lymph node and Peyer's patch), Kuffer cells, Langerhans cells, and bone marrow stromal cells. F4/80 has also been shown on a subset of dendritic cells. The biological ligand of F4/80 has not been identified, but it has been reported that F4/80 is required for induction of CD8+ T cells-mediated peripheral tolerance.

Other Names: EMR1, Ly71
Structure: EGF-TM7 family member, 160 kD glycoprotein
Distribution: Majority of tissue macrophages including peritoneal macrophages, macrophages in lung, gut, thymus and red pulp of spleen, Kuffer cells, Langerhans cells, bone marrow stromal cells, and a subset of dendritic cells
Function: Induction of immunological tolerance
Antigen References:

1. Austy JM and Gordon S. 1981. Eur. J. Immunol. 11:805.
2. Hume DA, et al. 1983. J. Exp. Med. 158:1522.
3. Ruedl C, et al. 1996. Eur. J. Immunol. 26:1801.
4. McKnight AJ, et al. 1996. J. Biol. Chem. 271:486.
5. Lin HH, et al. 2005. J. Exp. Med. 201:1615.