PE Mouse Anti-Human TCR Vβ5.1

Consider BD Horizon RealYellow™ 586 (RY586) Reagents, a bright and clean fluorochrome alternative to PE off the yellow-green laser. RY586 can be used alongside PE on spectral cytometers. More Info

BD Pharmingen™ PE Mouse Anti-Human TCR Vβ5.1

Name: PE Mouse Anti-Human TCR Vβ5.1
Brand: BD Pharmingen™
SKU: 567042

Clone LC4

(RUO)

Two-color flow cytometric analysis of TCR Vβ5.1 expression on human peripheral blood T lymphocytes. Human peripheral blood cells were stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 561811/555335/561810) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human TCR Vβ5.1 antibody (Cat. No. 567042/567043; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of TCR Vβ5.1 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Pharmingen™

Alternative Name: TCR V beta 5.1; TCR Vb5.1; TRBV5-1; TRBV51

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human T acute lymphoblastic leukemia Cell Line

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

RRID: AB_2870016

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: 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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

PE Mouse IgG1, κ Isotype Control RUO

Size 0.1 mg Cat No. 554680

Antibody Details

LC4

The LC4 monoclonal antibody specifically recognizes the human variable beta 5.1 (Vβ5.1) domain of the beta subunit for the αβ T cell receptor for antigen (TCR αβ). TCR Vβ5.1 is encoded by TRBV5-1 (T cell receptor beta variable 5-1), one of five functional genes within the TRBV5 subgroup in the T cell receptor beta (TRB) locus. The heterodimeric TCR αβ is composed of two disulfide-linked transmembrane glycoproteins, ie, highly variable TCRα and TCRβ chains. These chains are each comprised of an extracellular N-terminal variable (V) region domain followed by a constant (C) region domain, a transmembrane region, and a short C-terminal cytoplasmic tail. The TCR Vβ repertoire is known to be extensive due to the many different combinations of TCR gene segments (Vβ, Dβ, and Jβ) as well as junctional region diversity. TCR Vβ5.1 is variably expressed on subsets of TCR αβ-positive thymocytes and peripheral CD4+ or CD8+ T cells. In association with the CD3 complex of signaling proteins, the TCR αβ recognizes peptide-major histocompatibility complexes (pMHC) that are displayed on other cells to mediate cellular responses. The LC4 antibody is useful for analyzing the levels of TCR Vβ5.1 expressed by individual cells as well as the numbers or frequencies of TCR Vβ5.1-positive cells within test samples. The LC4 antibody can be used to help characterize the TCR Vβ repertoires of T cell populations during health as well as in response to vaccination, infectious disease, aging, transplantation, autoimmunity or cancer.

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

Citations & References

Development References (5)

Choi YW, Kotzin B, Lafferty J, et al. A method for production of antibodies to human T-cell receptor beta-chain variable regions.. Proc Natl Acad Sci USA. 1991; 88(19):8357-61. (Clone-specific: Flow cytometry). View Reference
Janson CH, Grunewald J, Osterborg A, et al. Predominant T cell receptor V gene usage in patients with abnormal clones of B cells.. Blood. 1991; 77(8):1776-80. (Clone-specific: Flow cytometry). View Reference
Maecker HT, Levy R. Prevalence of antigen receptor variants in human T cell lines and tumors.. J Immunol. 1989; 142(4):1395-404. (Immunogen: Flow cytometry, Functional assay, Immunocytochemistry). View Reference
van den Beemd R, Boor PP, van Lochem EG, et al. Flow cytometric analysis of the Vbeta repertoire in healthy controls.. Cytometry. 2000; 40(4):336-45. (Clone-specific: Flow cytometry). View Reference
von Bonin M, Wermke M, Cosgun KN, et al. In vivo expansion of co-transplanted T cells impacts on tumor re-initiating activity of human acute myeloid leukemia in NSG mice.. PLoS ONE. 2013; 8(4):e60680. (Clone-specific: Flow cytometry). View Reference

View All (5)

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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.