Biolegend, 123131, Brilliant Violet 421™ anti-mouse F4/80 Antibody, 125microl

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.
125microl
Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
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.
Concentration: µg sizes: 0.2 mg/mLµL sizes: lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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 - Verified SB - Reported in the literature, not verified in house
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 in 100 µl staining volume 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. Learn more about Brilliant Violet™. 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.
Excitation Laser: Violet Laser (405 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: Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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
Product Citations: Zahr A, et al. 2016. Nat Commun. 7:10363. PubMed Wang X, et al. 2019. Cell Res. 29:787. PubMed Yu X, et al. 2020. Nat Commun. 11:1110. PubMed Gao L, et al. 2023. J Nanobiotechnology. 21:56. PubMed Wu Q, et al. 2023. Front Immunol. 13:1081719. PubMed Swan SL, et al. 2023. Front Immunol. 14:1085547. PubMed Wilson NG, et al. 2023. iScience. 26:105991. PubMed Bautista CA, et al. 2023. Front Physiol. 14:1122348. PubMed Guo Z, et al. 2023. Int J Mol Sci. 24:. PubMed Zhou R, et al. 2022. EBioMedicine. 75:103762. PubMed Miranda K, et al. 2022. iScience. 25:104994. PubMed Wang L, et al. 2022. Cell Metab. 34:75. PubMed Zhang Y, et al. 2022. J Cereb Blood Flow Metab. 42:952. PubMed Kang H, et al. 2022. Adv Mater. 34:e2106500. PubMed Brog RA, et al. 2022. Cancer Immunol Res. 10:962. PubMed Liu Z, et al. 2022. Cancer Commun (Lond). 42:971. PubMed Zhang Z, et al. 2022. Mol Nutr Food Res. 66:e2200300. PubMed Lu Z, et al. 2022. Cell Rep. 41:111482. PubMed Siret C, et al. 2022. Nat Commun. 13:7366. PubMed Al-Saafeen BH, et al. 2023. Front Immunol. 13:1017780. PubMed Arimoto KI, et al. 2023. Nat Commun. 14:251. PubMed Taniguchi S, et al. 2023. Nat Commun. 14:143. PubMed Nettersheim FS, et al. 2023. Front Cardiovasc Med. 9:1076808. PubMed Yang L, et al. 2023. Sci Rep. 13:1827. PubMed Wu Q, et al. 2023. Cell Rep. 42:112057. PubMed Zhao X, et al. 2022. Cell Mol Immunol. 19:540. PubMed Hailemichael Y, et al. 2022. Cancer Cell. 40:509. PubMed Hu K, et al. 2022. J Extracell Vesicles. 11:e12199. PubMed Jaiswal AK, et al. 2020. Immunobiology. 225:151956. PubMed Lyons J, et al. 2018. PLoS Biol. 16:e2002417. PubMed Hong M, et al. 2021. Int J Mol Sci. 22:. PubMed Lefebvre MN, et al. 2021. Cell Rep. 37:109956. PubMed He J, et al. 2022. Front Immunol. 12:760138. PubMed Ma J, et al. 2020. Adv Sci (Weinh). 7:2000609. PubMed Mogilenko DA, et al. 2020. Immunity. 54(1):99-115.e12. PubMed Merz SF, et al. 2019. Nat Commun. 10:2312. PubMed Shen Y, et al. 2021. Comput Struct Biotechnol J. 19:5360. PubMed Xu ZH, et al. 2021. Mediators Inflamm. 2021:8856326. PubMed Feng J, et al. 2021. iScience. 24:103004. PubMed Iturri L, et al. 2021. Immunity. . PubMed Clement M, et al. 2016. PLoS Pathog. 12:e1006050. PubMed Sakai M, et al. 2020. Immunity. 51(4):655-670. PubMed Nakamoto N, et al. 2019. Nat Microbiol. 4:492. PubMed Li J, et al. 2021. Front Immunol. 12:649285. PubMed Fitzgerald AA, et al. 2021. J Immunother Cancer. 9:. PubMed Sun K, et al. 2016. J Exp Med. 213: 1851 - 1864. PubMed Laumonnier Y, et al. 2020. Curr Protoc Immunol. 130:e100. PubMed Brunner JS, et al. 2020. Nat Commun. 0.757638889. PubMed Gómez-Díaz C, et al. 2021. iScience. 24:103241. PubMed Sugimoto C, et al. 2022. Elife. 11:. PubMed Croasdell A, et al. 2016. J Immunol. 196: 2742 - 2752. PubMed Karsten C, et al. 2015. J Immunol. 194:1841. PubMed Lyons J, et al. 2018. Sci Signal. 11. PubMed Zhang X, et al. 2022. Pharmaceutics. 14:. PubMed Liu Y, et al. 2022. Nat Commun. 13:2665. PubMed Glass A, et al. 2013. J Immunol. 190:4830. PubMed Müller P, et al. 2015. Sci Transl Med. 7: 315ra188. PubMed Doty DT, et al. 2020. Int J Mol Sci. 21:00. PubMed Sokol CL et al. 2018. Immunity. 49(3):449-463 . PubMed Kobayashi S, et al. 2019. J Immunol. 203:1447. PubMed Samarchith P Kurup et al. 2019. Cell host & microbe. 25(4):565-577 . PubMed Palathingal Bava E, et al. 2022. JCI Insight. 7:. PubMed Tu J, et al. 2022. Theranostics. 12:747. PubMed Du W, et al. 2021. Mol Cancer Res. 19:1412. PubMed Opzoomer JW, et al. 2021. Sci Adv. 7:eabg9518. PubMed Paterson N, et al. 2022. Elife. 11:. PubMed Topper MJ et al. 2017. Cell. 171(6):1284-1300 . PubMed Mrdjen D et al. 2018. Immunity. 48(2):380-395 . PubMed Eggold JT, et al. 2022. Mol Cancer Ther. 21:371. PubMed Zenker S, et al. 2014. J Immunol. 192:2830. PubMed Li J, et al. 2020. Elife. 9:00. PubMed Schadt L, et al. 2020. Cell Reports. 29(5):1236-1248.e7.. PubMed Fennell LM, et al. 2020. EMBO J. 39:e103303. PubMed Chi A, et al. 2022. Front Endocrinol (Lausanne). 13:871548. PubMed Friedman DJ, et al. 2021. Cancer Immunol Res. 9:952. PubMed Guo P, et al. 2021. J Immunol. 207:408. PubMed Mulas F, et al. 2020. Cell Mol Immunol. . PubMed Cao DY, et al. 2020. J Biol Chem. 295:1369. PubMed Textor A, et al. 2014. Cancer Res. 74:6769. PubMed Kumar MP, et al. 2018. Cell Rep. 25:1458. PubMed Bergin SM, et al. 2021. Brain Behav Immun. 95:477. PubMed Enders M, et al. 2020. J Immunol. 204:87. PubMed Engler AE, et al. 2020. Cell Reports. 33(13):108553. PubMed He J, et al. 2019. J Clin Invest. 130. PubMed Alikhanyan K, et al. 2020. Immun Inflamm Dis. 8:181. PubMed Troutman TD, et al. 2021. STAR Protocols. 2(1):100363. PubMed Denny JE, et al. 2019. Sci Rep. 2.786111111. PubMed Yu X, et al. 2019. Nat Commun. 10:574. PubMed Khare P, et al. 2017. J Autoimmun. 10.1016/j.jaut.2017.09.002. PubMed Becker W, et al. 2021. J Crohns Colitis. 15:1032. PubMed Sun M, et al. 2021. J Immunother Cancer. 9:. PubMed Wei Z, et al. 2021. Nat Commun. 0.805555556. PubMed Chuang CH, et al. 2020. Cancer Research. 81:567. PubMed Lim J et al. 2019. Elife. 8 pii: e44452. PubMed McDonald LT, et al. 2018. Am J Physiol Heart Circ Physiol. 315:H92. PubMed Guan X, et al. 2022. Nat Commun. 13:2834. PubMed Babagana M, et al. 2021. Aging (Albany NY). 13:19207. PubMed Ryan NM, et al. 2022. Front Immunol. 13:932742. PubMed Nahrendorf W, et al. 2021. eLife. 10:00. PubMed Muri J, et al. 2020. eLife. 9:e53627.. PubMed Lu Y, et al. 2020. Immunity. 52:782. PubMed Chan WY, et al. 2019. Infect Immun. 87:. PubMed Matsumura T, et al. 2022. Nat Commun. 13:7064. PubMed Da Mesquita S, et al. 2021. Science Advances. 7(21):. PubMed Ledo JH, et al. 2020. PLoS One. e0237773:15. PubMed Barbet G, et al. 2018. Immunity. 48:584. PubMed Bagati A, et al. 2020. Cancer Cell. 39(1):54-67.e9. PubMed Suzuki T, et al. 2017. Cell Rep. 18(8):2045-2057. PubMed Muri J, et al. 2020. Cell Reports. 29(9):2731-2744.e4.. PubMed Heinen A, et al. 2019. Mol Ther. 27:46:00. PubMed Clemente–Casares X, et al. 2017. Immunity. 47:974. PubMed Hobbs BE, et al. 2021. Pathogens. 10:. PubMed Alhudaithi SS, et al. 2020. Mol Pharm. 17:4691. PubMed Crichton M, et al. 2016. Sci Rep. 6:27217. PubMed Liao X, et al. 2022. JCI Insight. 7:. PubMed Mujal AM, et al. 2022. Cancer Immunol Res. 10:403. PubMed Wang Y, et al. 2021. Nat Commun. 0.570833333. PubMed Silva HM, et al. 2019. J Exp Med. 216:786. PubMed Lee C, et al. 2020. Front Immunol. 11:77. PubMed Säwen P et al. 2018. eLife. 7 pii: e41258. PubMed Fan HH, et al. 2021. Methods Mol Biol. 2276:203. PubMed Zhang Y, et al. 2021. JCI Insight. 6:e150735. PubMed Liu X, et al. 2021. Adv Sci (Weinh). 8:e2100233. PubMed Jaeger N, et al. 2020. Cell Rep. 33:108331. PubMed Shan Z, et al. 2021. eLife. 10:00. PubMed Li Z et al. 2018. Immunity. 49(4):640-653 . PubMed Ying L, et al. 2021. Front Cell Dev Biol. 9:672032. PubMed Hsu HP, et al. 2021. J Biol Chem. 296:100419. PubMed Schuster EM, et al. 2022. Nat Metab. 4:856. PubMed Ledo JH, et al. 2020. Mol Psychiatry. . PubMed Cai W, et al. 2019. J Neuroinflammation. 0.788194444. PubMed Reinke S, et al. 2020. Cell Reports. 30(8):2501-2511. PubMed Pessoa Rodrigues C, et al. 2020. Sci Adv. 6:eaaz4815. PubMed Tong Y, et al. 2018. EBioMedicine. 39:132. PubMed Yao W, et al. 2017. EBioMedicine. 10.1016/j.ebiom.2017.07.014. PubMed
RRID: AB_10901171 (BioLegend Cat. No. 123131) AB_2563102 (BioLegend Cat. No. 123137) AB_11203717 (BioLegend Cat. No. 123132)
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: 13733
UniProt: View information about F4/80 on UniProt.org
Clone: BM8
Regulatory Status: RUO
Other Names: EMR1, Ly71
Isotype: Rat IgG2a, κ
Q: What is the F/P ratio range of our BV421™ format antibody reagents?
A: It is lot-specific. On average it ranges between 2-4.
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Q: Are other fluorophores compatible with IBEX?
A: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
Q: The same antibody works in one tissue type but not another. What is happening?
A: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.