BD Horizon™ RB613 Mouse Anti-Human CD272 (BTLA)
Clone J168-540 (RUO)
Multiparameter flow cytometric analysis of CD272 (BTLA) expression on Human peripheral blood leukocytes. Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426; Top Plots) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plots) or BD Horizon™ RB613 Mouse Anti-Human CD272 (BTLA) antibody (Cat. No. 571259/571319; Right Plots). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of CD272 (BTLA) [or Ig Isotype control staining] versus CD3 (Top Plots) or side light-scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter signals of intact leukocyte populations. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of CD272 (BTLA) expression on Human peripheral blood leukocytes. Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426; Top Plots) and with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plots) or BD Horizon™ RB613 Mouse Anti-Human CD272 (BTLA) antibody (Cat. No. 571259/571319; Right Plots). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of CD272 (BTLA) [or Ig Isotype control staining] versus CD3 (Top Plots) or side light-scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter signals of intact leukocyte populations. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
BTLA1; B- and T-lymphocyte-associated protein
Human (QC Testing)
Mouse IgG1, κ
Human BTLA Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl/test
IX 47
151888
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB613 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571106
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
BV421 Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
562426
571259 Rev. 1
Antibody Details
J168-540
The J168-540 monoclonal antibody specifically binds to human CD272, also known as BTLA (B and T lymphocyte attenuator), an inhibitory receptor expressed in bone marrow and thymus on developing B and T cells. In the periphery, BTLA is expressed by B cells, T cells, bone marrow-derived dendritic cells, and macrophages. After T cell activation, BTLA appears to be expressed at higher levels on Th1 cell populations than in Th2 cell populations. Upon binding the herpesvirus entry mediator (HVEM/LIGHT-R/CD270), CD272 undergoes tyrosine phosphorylation and inhibits T cell proliferation in a BTLA-dependent manner. CD272 has structural similarities to two other lymphocyte inhibitory receptors, CTLA-4 and PD-1 and is a member of the CD28-like family of coreceptors. Based on these observations, CD272 is considered to be a negative regulator of lymphocyte activation and/or function.
571259 Rev. 1
Format Details
RB613
The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
RB613
Blue 488 nm
492 nm
613 nm
571259 Rev.1
Citations & References
View product citations for antibody "571259" on CiteAb
Development References (5)
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Compaan DM, Gonzalez LC, Tom I, Loyet KM, Eaton D, Hymowitz SG. Attenuating lymphocyte activity: the crystal structure of the BTLA-HVEM complex. J Biol Chem. 2005; 280(47):39553-39561. (Biology). View Reference
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Hobo W, Norde WJ, Schaap N, et al. B and T lymphocyte attenuator mediates inhibition of tumor-reactive CD8+ T cells in patients after allogeneic stem cell transplantation. J Immunol. 2012; 189(1):39-49. (Clone-specific: Flow cytometry). View Reference
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Llinas L, Lazaro A, de Salort J, Matesanz-Isabel J, Sintes J, Engel P. Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry. Immunol Lett. 2011; 134(2):113-121. (Clone-specific: Flow cytometry). View Reference
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Sedy JR, Gavrieli M, Potter KG, et al. B and T lymphocyte attenuator regulates T cell activation through interaction with herpesvirus entry mediator. Nat Immunol. 2005; 6(1):90-98. (Biology). View Reference
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Watanabe N, Gavrieli M, Sedy JR, et al. BTLA is a lymphocyte inhibitory receptor with similarities to CTLA-4 and PD-1. Nat Immunol. 2003; 4(7):670-679. (Biology). View Reference
571259 Rev. 1
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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.