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RB705 Mouse Anti-Mouse CD272 (BTLA)
Product Details
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BD OptiBuild™
B- and T-lymphocyte attenuator; Btla
Mouse (Tested in Development)
Mouse IgG1, κ
C57BL/6 BTLA Ig Domain Protein
Flow cytometry (Qualified)
0.2 mg/ml
208154
Aqueous buffered solution containing ≤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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  7. 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.
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. 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.
757563 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.

757563 Rev. 1
Format Details
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RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
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RB705
Blue 488 nm
498 nm
707 nm
757563 Rev.1
Citations & References
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View product citations for antibody "757563" 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
757563 Rev. 1

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.