Biolegend, 285165, Spark Blue™ 574 anti-mouse F4/80 (Flexi-Fluor™) Antibody, 50μg

F4/80, also known as EMR1 or Ly71, is a 160 kD glycoprotein of the epidermal growth factor (EGF)-transmembrane 7 (TM7) family. F4/80 has been widely used as a murine macrophage marker. It is expressed on a majority of tissue macrophages, including macrophages in the lung, gut, peritoneal cavity, thymus, and red pulp of the spleen, Kupffer cells, Langerhans cells, microglia, and certain dendritic cells. It is not expressed on macrophages located in the T cell areas of the spleen, lymph node, or Peyer's patch. The biological ligand of F4/80 has not been identified, but it has been reported that F4/80 is required for the induction of CD8 + T cells-mediated peripheral tolerance.
50μg
Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: Murine macrophages
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.
Concentration: 0.2 mg/mL
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
Recommended Usage: Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. * Spark Blue™ 574 has a maximum excitation of 506 nm and a maximum emission of 574 nm.
Excitation Laser: Blue Laser (488 nm)
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, Western blotting, and spatial biology (IBEX)12,13.
Additional Product Notes: For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.
Application References(PubMed link indicates BioLegend citation): Schaller E, et al. 2002. Mol. Cell. Biol. 22:8035. (IHC) Stevceva L, et al. 2001. BMC Clin Pathol. 1:3. (IHC) Kobayashi M, et al.2008. J. Leukoc. Biol. 83:1354. PubMed Poeckel D, et al. 2009. J. Biol Chem. 284:21077. PubMed Glass AM, et al. 2013. J. Immunol. 190:4830. PubMed Koehm S, et al. 2007. J. Allergy Clin. Immunol. 120:570. (IHC) Rankin AL, et al. 2010. J. Immunol. 184:1526. (IHC) Sasi SP, et al. 2014. J Biol Chem. 289:14178. PubMed Thakus VS, et al. 2014. Toxicol Lett. 230:322. PubMed Watson NB, et al. 2015. J Immunol. 194:2796. PubMed Hirakawa H, et al. 2015. PLoS One. 10:119360. PubMed Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID: AB_3662505 (BioLegend Cat. No. 285165)
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, Kupffer cells, Langerhans cells, bone marrow stromal cells, and a subset of dendritic cells
Function: Induction of immunological tolerance
Cell Type: Dendritic cells, Langerhans cells, Macrophages, Tregs
Biology Area: Cell Biology, Immunology, Innate Immunity, Neuroinflammation, Neuroscience
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.
Gene ID: NA
UniProt: View information about F4/80 on UniProt.org
Clone: BM8
Regulatory Status: RUO
Other Names: EMR1, Ly71
Isotype: Rat IgG2a, κ
Q: What are Flexi-Fluors?
A: Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer.
Q: How quickly will I receive my order?
A: We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.
Q: How are Flexi-Fluors different from regular catalog products?
A: Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.
Q: How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?
A: Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
Q: I can’t find the antibody-dye combination that I need. When will it be available?
A: We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services.
Q: I need help to validate the performance of my Flexi-Fluor. Who should I contact?
A: Please get in touch with Technical Service for assistance.
Q: Can I order more than 50 μg of a Flexi-Fluor?
A: Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.
Q: What is the expiration date of my Flexi-Fluor?
A: Expiration dates can be found on the vial label or by using our CoA lookup tool.
Q: What if my requested antibody-dye combination doesn't perform as expected?
A: Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.