Biolegend, 303105, PE anti-human CD31 Antibody, 25tests

CD31 is a 130-140 kD type I transmembrane glycoprotein also known as platelet endothelial cell adhesion molecule-1 (PECAM-1) or Endocam. It is expressed on monocytes, platelets, granulocytes, endothelial cells and lymphocyte subsets. CD31 has been reported to bind CD38 and be involved in wound healing, angiogenesis, and cellular migration in an inflammatory situation.
25tests
Verified Reactivity: Human, Cynomolgus, Rhesus
Reported Reactivity: African Green, Baboon
Antibody Type: Monoclonal
Host Species: Mouse
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 PE 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: FC - Quality tested 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 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.
Excitation Laser: Blue Laser (488 nm)Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes: Clone WM59 has been reported to recognize the D2 extracellular portion of CD31.Additional reported applications (for the relevant formats) include: immunofluorescence microscopy2, immunohistochemical staining of acetone-fixed frozen tissue sections8, blocking of platelet aggregation3, and spatial biology (IBEX)11,12. Clone WM59 is not recommended for immunohistochemical staining of formalin-fixed paraffin-embedded sections. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 303143 & 303144).The purified WM59 antibody is useful as a capture antibody for a sandwich ELISA assay, when used in conjunction with biotin anti-human CD31 antibody (Cat. No. 536604) antibody as the detection antibody.
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): Schlossman S, et al. Eds. 1995. Leucocyte Typing V Oxford University Press. New York. Muczynski KA, et al. 2003. J. Am. Soc. Nephrol. 14:1336. (IF) Wu XW, et al. 1997. Arterioscl. Throm. Vas. 17:3154. (Block) Nagano M, et al. 2007. Blood 110:151. (FC) PubMed MacFadyen JR, et al. 2005. FEBS Lett. 579:2569. PubMed Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Sestak K, et al. 2007. Vet. Immunol. Immunopathol. 119:21. Wicki A, et al. 2012. Clin. Cancer Res. 18:454. (FC, IHC) PubMed Oeztuerk-Winder F, et al. 2012. EMBO J. 31:3431. (FC) PubMed Bushway ME, et al. 2014. Biol Reprod. 90(5): 110 (IF) PubMed Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations: Xin Y, et al. 2021. Stem Cell Res Ther. 49:12. PubMed Zhu H, et al. 2022. Cell Transplant. 31:9636897221106996. PubMed Pal D, et al. 2023. Nat Commun. 14:1129. PubMed Liu Y, et al. 2023. Int J Stem Cells. . PubMed Liu C, et al. 2022. Regen Ther. 21:192. PubMed Raggi C, et al. 2022. Curr Protoc. 2:e389. PubMed Robb KP, et al. 2022. Front Immunol. 13:972095. PubMed Guahmich NL, et al. 2023. Commun Biol. 6:7. PubMed Lu T, et al. 2023. Cancer Immunol Immunother. :. PubMed Tuohinto K, et al. 2023. PLoS Pathog. 19:e1010753. PubMed Iio K, et al. 2023. Immun Ageing. 20:8. PubMed Gao Q, et al. 2019. J Exp Med. 216:688. PubMed Gong Y, et al. 2021. Aging (Albany NY). 13:20629. PubMed Lu D, et al. 2022. Dis Markers. 2022:3556372. PubMed Zhang J, et al. 2022. Int J Mol Sci. 23:. PubMed Pettinato G, et al. 2019. Sci Rep. 9:8920. PubMed Yu C, et al. 2020. Sci Rep. 10:14521. PubMed Witkowski MT, et al. 2020. Cancer Cell. 37:867. PubMed Alhaj Hussen K, et al. 2017. Immunity. 47:680. PubMed Revel-Vilk S, et al. 2015. Clin Immunol. 159: 84-92. PubMed Nagano M, et al. 2007. Blood . 110:151. PubMed Nagai N, et al. 2018. PLoS Genet. 14:e1007826. PubMed Xu P, et al. 2020. Cancer Immunol Res. 8:1193. PubMed Zhang Y, et al. 2018. Stem Cells Int. 2018:7159465. PubMed Dedeoglu B, et al. 2016. PLoS One. 11: 0150826. PubMed Scherpenisse M, et al. 2021. MBio. 12:. PubMed Iio K, et al. 2019. Sci Rep. 9:813. PubMed Sun K, et al. 2022. Nat Commun. 13:4943. PubMed Pan C, et al. 2019. Int J Mol Med. 44:1629. PubMed Kerdidani D, et al. 2022. J Exp Med. 219:. PubMed Dong N, et al. 2022. World J Stem Cells. 14:556. PubMed Wei X, et al. 2019. Int J Mol Med. 44:1425. PubMed Künzel K, et al. 2022. Heliyon. 8:e10365. PubMed Khanh VC, et al. 2021. Stem Cells Dev. 30:758. PubMed
RRID: AB_314331 (BioLegend Cat. No. 303105) AB_314332 (BioLegend Cat. No. 303106)
Structure: Ig superfamily, type I transmembrane glycoprotein, 130-140 kD
Distribution: Monocytes, platelets, granulocytes, endothelial cells, lymphocyte subset
Function: Cell adhesion, signal transduction
Ligand/Receptor: CD38
Cell Type: Endothelial cells, Granulocytes, Lymphocytes, Monocytes, Neutrophils, Platelets
Biology Area: Angiogenesis, Cell Adhesion, Cell Biology, Immunology, Neuroinflammation, Neuroscience
Molecular Family: Adhesion Molecules, CD Molecules
Antigen References: DeLisser H, et al. 1994. Immunol. Today 15:490. Newman P, 1997. J. Clin. Invest. 99:3. Fawcett J, et al. 1995. J. Cell Biol. 128:1229.
Gene ID: 5175
UniProt: View information about CD31 on UniProt.org
Clone: WM59
Regulatory Status: RUO
Workshop: V P025
Other Names: PECAM-1, EndoCAM
Isotype: Mouse IgG1, κ
Q: What type of PE do you use in your conjugates?
A: We use R-PE in our conjugates.
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.