Flow cytometric analysis of CD62P expression on mouse platelets. Mouse platelets were either not activated (Left Panel) or activated with human thrombin (Sigma T-8885; 20 U/ml final concentration; 5 min at 37°C; Right Panel). The platelets were stained with either BD Horizon™ BV421 Rat IgG1, λ Isotype Control (Cat. No. 562604; dashed line histograms) or BD Horizon™ BV421 Rat Anti-Mouse CD62P antibody (Cat. No. 564289; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of platelets. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD62P expression on mouse platelets. Mouse platelets were either not activated (Left Panel) or activated with human thrombin (Sigma T-8885; 20 U/ml final concentration; 5 min at 37°C; Right Panel). The platelets were stained with either BD Horizon™ BV421 Rat IgG1, λ Isotype Control (Cat. No. 562604; dashed line histograms) or BD Horizon™ BV421 Rat Anti-Mouse CD62P antibody (Cat. No. 564289; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of platelets. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Horizon™
P-selectin; Selp; LECAM3; LYAM3; PADGEM; GMP-140; Grmp
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG1, λ
P-selectin-IgG1 Fusion
Flow cytometry (Routinely Tested)
0.2 mg/ml
20344
AB_2738732
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
BV421 Rat IgG1, λ Isotype Control RUO
Size
50 µg
Cat No.
562604
564289 Rev. 1
Antibody Details
RB40.34
The RB40.34 monoclonal antibody specifically binds to mouse P-selectin (CD62P), a 140 kDa protein which is expressed on activated platelets, activated endothelial cells, and megakaryocytes. P-selectin mediates the adhesion of neutrophils and monocytes to activated platelets and endothelial cells, mediates leukocyte rolling, and is involved in the migration of leukocytes into inflamed tissues. CD24 and CD162 (PSGL-1) are ligands of CD62P. mAb RB40.34 can block mouse P-selectin binding to its ligands in vitro and in vivo.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
564289 Rev. 1
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
Violet 405 nm
407 nm
423 nm
564289 Rev.1
Citations & References
Development References (9)
-
Aigner S, Ruppert M, Hubbe M, et al. Heat stable antigen (mouse CD24) supports myeloid cell binding to endothelial and platelet P-selectin. Int Immunol. 1995; 7(10):1557-1565. (Biology). View Reference
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Austrup F, Vestweber D, Borges E, et al. P- and E-selectin mediate recruitment of T-helper-1 but not T-helper-2 cells into inflammed tissues. Nature. 1997; 385(6611):81-83. (Clone-specific: Blocking). View Reference
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Bosse R, Vestweber D. Only simultaneous blocking of the L- and P-selectin completely inhibits neutrophil migration into mouse peritoneum. Eur J Immunol. 1994; 24(12):3019-3024. (Immunogen: Blocking, ELISA, Flow cytometry, Immunoprecipitation). View Reference
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Hirata T, Furie BC, Furie B. P-, E-, and L-selectin mediate migration of activated CD8+ T lymphocytes into inflamed skin. J Immunol. 2002; 169(8):4307-4313. (Clone-specific: Blocking). View Reference
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Katakai T, Mori KJ, Masuda T, Shimizu A. Selective accumulation of type 1 effector cells expressing P-selectin ligand and/or alpha(4)beta(7)-integrin at the lesions of autoimmune gastritis. Int Immunol. 2002; 14(2):167-175. (Clone-specific: Immunohistochemistry). View Reference
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Ley K, Bullard DC, Arbones ML, et al. Sequential contribution of L- and P-selectin to leukocyte rolling in vivo. J Exp Med. 1995; 181(2):669-675. (Clone-specific: Blocking). View Reference
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Pendl GG, Robert C, Steinert M, et al. Immature mouse dendritic cells enter inflamed tissue, a process that requires E- and P-selectin, but not P-selectin glycoprotein ligand 1. Blood. 2002; 99(3):946-956. (Clone-specific: Blocking). View Reference
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Tietz W, Allemand Y, Borges E, et al. CD4+ T cells migrate into inflamed skin only if they express ligands for E- and P-selectin. J Immunol. 1998; 161(2):963-970. (Clone-specific: Blocking). View Reference
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Yang J, Galipeau J, Kozak CA, Furie BC, Furie B. Mouse P-selectin glycoprotein ligand-1: molecular cloning, chromosomal localization, and expression of a functional P-selectin receptor. Blood. 1996; 87(10):4176-4186. (Biology). View Reference
564289 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
