Biolegend, 141717, Brilliant Violet 421™ anti-mouse CD206 (MMR) Antibody, 125microl

CD206, also known as mannose receptor (MR), is a 175 kD type I membrane protein. It is a pattern recognition receptor (PRR) belonging to the C-type lectin superfamily. MR is expressed on macrophages, dendritic cells, Langerhans cells, and hepatic or lymphatic endothelial cells. MR recognizes a range of microbial carbohydrates bearing mannose, fucose, or N-acetyl glucosamine through its C-type lectin-like carbohydrate recognition domains, sulfated carbohydrate antigens through its cysteine-rich domain, and collagens through its fibronectin type II domain. MR mediates endocytosis and phagocytosis as well as activation of macrophages and antigen presentation. It plays an important role in host defense and provides a link between innate and adaptive immunity. Recently, MR on lymphatic endothelial cells was found to be involved in leukocyte trafficking and a contributor to the metastatic behavior of cancer cells. It suggests that MR may be a potential target in controlling inflammation and cancer metastasis by targeting the lymphatic vasculature.
125microl
Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: Recombinant mouse CD206 (MMR)
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: 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: ICFC - Quality tested FC, IHC-F - Verified SB - Reported in the literature, not verified in house
Recommended Usage: Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, 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 immunohistochemistry on frozen tissue sections, a concentration range of 2.5 - 5.0 µg/mL is suggested. 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: Clone C068C2 recognizes a region similar to clone MR5D3, based on the ability of the clones to block each other. Additional reported applications (for the relevant formats) include: spatial biology (IBEX)4,5.
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): Keller J, et al. 2012. Biochem Biophys Res Commun. 417:217. PubMed Ito H, et al. 2012. J Am Soc Nephrol. 23:1797. PubMed Yang X, et al. 2015. PNAS. 112:2900. 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: Gonzalez C, et al. 2022. Life Sci. 309:121003. PubMed Lu L, et al. 2023. Adv Sci (Weinh). 10:e2206212. PubMed Stratoulias V, et al. 2023. Nat Neurosci. 26:1008. PubMed Yu Y, et al. 2023. Cancer Cell Int. 23:92. PubMed Brigo N, et al. 2023. Cells. 12:. PubMed Liu M, et al. 2022. Cells. 11: . PubMed Hattori Y, et al. 2023. Cell Rep. 42:112092. PubMed Ma Y, et al. 2022. Mol Biol Rep. 49:3975. PubMed Gomez-Salinero JM, et al. 2022. Cell Stem Cell. 29:593. PubMed Kim SS, et al. 2022. Cancers (Basel). 14:. PubMed Trofimova O, et al. 2021. Vaccines (Basel). 9:. PubMed Conforti A, et al. 2021. PLoS One. 16:e0259894. PubMed Vicario N, et al. 2021. Cell Death Disease. 12(7):625. PubMed Nguyen CM, et al. 2019. Diabetes. 68:1499. PubMed Simonneau M, et al. 2018. Oncotarget. 9:36457. PubMed Fulham MA, et al. 2019. Am J Physiol Cell Physiol. 317:C687. PubMed Ayoub M, et al. 2019. Sci Rep. 9:6348. PubMed Das LM, et al. 2019. Autophagy. 15:813. PubMed Kimbrough D, et al. 2018. J Mol Cell Cardiol. 119:51. PubMed Hanasoge Somasundara AV, et al. 2021. Cell Rep. 37:110099. PubMed Cho YK, et al. 2022. Nat Commun. 13:4084. PubMed Yang P, et al. 2022. Nat Commun. 13:5782. PubMed Macdougall CE et al. 2018. Cell metabolism. 27(3):588-601 . PubMed Giampazolias E, et al. 2021. Cell. . PubMed Alam Z, et al. 2020. Cell Rep. 107825:31. PubMed Xiao Y, et al. 2022. Nat Commun. 13:758. PubMed Parigiani MA, et al. 2020. Cancers (Basel). 12:00. PubMed Ito F, et al. 2015. PLoS One. 10: 0143370. PubMed Baumann D, et al. 2020. Nat Commun. 1.969444444. PubMed Yang N, et al. 2021. NPJ Precis Oncol. 5:37. PubMed Chmielewski M and Abken H 2017. Cell Rep.. 10.1016/j.celrep.2017.11.063. PubMed
RRID: AB_2562232 (BioLegend Cat. No. 141717)
Structure: Type I transmembrane protein, 175 kD, C-type lectin superfamily
Distribution: Macrophages, dendritic cells, Langerhans cells, liver endothelial cells
Function: Pathogen recognition, endocytosis and phagocytosis, antigen presentation
Ligand/Receptor: Antigen containing mannose, fucose, or an N-acetyl glucosamine
Cell Type: Dendritic cells, Endothelial cells, Langerhans cells, Macrophages
Biology Area: Cell Biology, Immunology, Innate Immunity, Signal Transduction
Molecular Family: CD Molecules
Antigen References: 1. Wileman TE, et al. 1986. P. Natl. Acad. Sci. USA 83:2501. 2. Apostolopoulos V, et al. 2001. Curr. Mol. Med. 1:469. 3. Burgdorf S, et al. 2006. J. Immunol. 176:6770. 4. McKenzie EJ, et al. 2007. J. Immunol. 178:4975.
Gene ID: 17533
UniProt: View information about CD206 on UniProt.org
Clone: C068C2
Regulatory Status: RUO
Other Names: MMR (macrophage mannose receptor), MR (mannose receptor), MRC1
Isotype: Rat IgG2a, κ
Q: Why is mouse CD206 stained intracellularly and not via surface staining?
A: Typically, mouse CD206 surface level is relatively low under normal conditions and so intracellular staining protocol is required to get better signal.
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.