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PE Mouse Anti-Mouse CD272 (BTLA)
PE Mouse Anti-Mouse CD272 (BTLA)
Two-color flow cytometric analysis of CD272 (BTLA) expression on Mouse splenic leucocytes. BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092) and with either PE Mouse IgG1, K Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse CD272 (BTLA) antibody (Cat. No. 568781/568782; Right Plot) at 0.25 µg /test. The bivariate pseudocolor density plot showing the correlated expression of CD272 (BTLA) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD272 (BTLA) expression on Mouse splenic leucocytes. BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092) and with either PE Mouse IgG1, K Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse CD272 (BTLA) antibody (Cat. No. 568781/568782; Right Plot) at 0.25 µg /test. The bivariate pseudocolor density plot showing the correlated expression of CD272 (BTLA) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
B- and T-lymphocyte attenuator; Btla
Mouse (QC Testing)
Mouse IgG1, κ
C57BL/6 BTLA Ig Domain Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
568781 Rev. 1
Antibody Details
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6F7/BTLA

The 6F7 monoclonal antibody specifically recognizes CD272 which is also known as B- and T-lymphocyte attenuator (BTLA) and is exclusively expressed on lymphoid cells. CD272 (BTLA) is a type 1 transmembrane glycoprotein that is encoded by Btla (B and T lymphocyte associated). CD272 (BTLA) contains a V-type Ig-like domain in its extracellular region followed by a transmembrane sequence, and a cytoplasmic domain with three tyrosine-based motifs, two immunoreceptor tyrosine-based inhibitory motifs (ITIM) and a Grb-2 recognition consensus sequence. The existence of three distinct BTLA alleles has been reported which encode molecules with different Ig domain structure and expression patterns on lymphoid cell subsets amongst different mouse strains. For example, whereas C57BL/6 and BALB/c mice both variably express CD272 (BTLA) on developing and mature T and B lymphocytes and dendritic cells (DC), C57BL/6 mice, but not BALB/c mice, also express CD272 (BTLA) on NK cells and macrophages. CD272 (BTLA) expression is upregulated by activated T cells including Th1, Th2, and anergic T cells. Herpesvirus entry mediator (HVEM), also known as CD270 and LIGHT-R, has been identified as a ligand for CD272 (BTLA). The crosslinking of CD272 (BTLA) by HVEM inhibits T-cell proliferation and cytokine production. CD272 (BTLA) is structurally like other coinhibitory receptors including CD152/CTLA-4 and CD279/PD-1. Although these coinhibitory receptors and their ligands maintain immunological homeostasis and self-tolerance, they may also serve as immune checkpoint molecules that inhibit adaptive immune responses against tumors and chronic infections.

568781 Rev. 1
Format Details
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PE
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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
568781 Rev.1
Citations & References
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View product citations for antibody "568781" on CiteAb

Development References (7)

  1. Crawford A, Wherry EJ. Editorial: Therapeutic potential of targeting BTLA.. J Leukoc Biol. 2009; 86(1):5-8. (Clone-specific). View Reference
  2. Han P, Goularte OD, Rufner K, Wilkinson B, Kaye J. An inhibitory Ig superfamily protein expressed by lymphocytes and APCs is also an early marker of thymocyte positive selection. J Immunol. 2004; 172(10):5931-5939. (Biology). View Reference
  3. Hurchla MA, Sedy JR, Gavrieli M, Drake CG, Murphy TL, Murphy KM. B and T lymphocyte attenuator exhibits structural and expression and is highly induced in anergic CD4+T cells. J Immunol. 2005; 174(6):3377-3385. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
  4. Nurieva RI, Chung Y, Martinez GJ, et al. Bcl6 mediates the development of T follicular helper cells. Science. 2009; 325(5943):1001-1005. (Biology). View Reference
  5. 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
  6. 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
  7. Watanabe N, Nakajima H. Coinhibitory molecules in autoimmune diseases. Clin Dev Immunol. 2012; 2012:1-7. (Biology). View Reference
View All (7) View Less
568781 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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.