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BV421 Rat Anti-Mouse CD8b

BD Horizon™ BV421 Rat Anti-Mouse CD8b

Clone YTS156.7.7.rMAb (also known as YTS156.7.7; YTS156)

(RUO)
BV421 Rat Anti-Mouse CD8b
Multicolor flow cytometric analysis of CD8b expression on viable Mouse splenic lymphocytes.  BALB/c Mouse splenic leukocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody (Cat. No. 563565) and with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; Left Plot) or BD Horizon™ BV421 Rat Anti-Mouse CD8b antibody (Cat. No. 569871/569947; Right Plot) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD8b (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. 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.
Multicolor flow cytometric analysis of CD8b expression on viable Mouse splenic lymphocytes.  BALB/c Mouse splenic leukocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody (Cat. No. 563565) and with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; Left Plot) or BD Horizon™ BV421 Rat Anti-Mouse CD8b antibody (Cat. No. 569871/569947; Right Plot) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD8b (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. 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 Horizon™
Cd8b1; Cd8b; Ly-3; Lyt-3; lymphocyte antigen 3
Mouse (QC Testing)
Rat IgG1, κ
Mouse Thymocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
12526
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 unconjugated antibody and free dye were 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.

   For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For U.S. patents that may apply, see bd.com/patents.
569871 Rev. 1
Antibody Details
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YTS156.7.7.rMAb

The YTS156.7.7.rMAb is a recombinant monoclonal antibody derived from YTS156.7.7 hybridoma cells that specifically recognizes the β chain of the CD8 differentiation antigen (CD8b), which is also known as Lymphocyte antigen 3 (Ly-3) or Lyt-3. CD8b is a single-pass type I transmembrane glycoprotein that is encoded by Cd8b1 (CD8 antigen, beta chain 1) which belongs to the Ig superfamily. CD8b is comprised of an N-terminal IgV domain in its extracellular region followed by a transmembrane region and a cytoplasmic tail. CD8b can bind to the CD8 α chain (CD8a) to form the disulfide-linked CD8 αβ (CD8ab) heterodimer also known as Ly2,3 or Lyt-2,3. This heterodimer is expressed on most thymocytes and a subpopulation of MHC class I-restricted mature T cells. CD8 αβ functions as an antigen coreceptor on the T-cell surface which interacts with MHC class I molecules on antigen-presenting cells. It participates in T-cell activation through its association with the T-cell receptor complex and the protein tyrosine kinase, lck (p56lck).

569871 Rev. 1
Format Details
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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.
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BV421
Violet 405 nm
407 nm
423 nm
569871 Rev.1
Citations & References
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Development References (5)

  1. Cobbold S, Martin G, Waldmann H. Monoclonal antibodies for the prevention of graft-versus-host disease and marrow graft rejection. The depletion of T cell subsets in vitro and in vivo.. Transplantation. 1986; 42(3):239-47. (Immunogen: Flow cytometry). View Reference
  2. Liunggren G, Liunggren HG, Dalianis T. T cell subsets involved in immunity against polyoma virus-induced tumors.. Virology. 1994; 198(2):714-6. (Clone-specific: In vivo exacerbation). View Reference
  3. Monney L1, Sabatos CA, Gaglia JL, et al. Th1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease.. Nature. 2002; 415(6871):536-541. (Clone-specific: Flow cytometry). View Reference
  4. Shore DA, Issafras H, Landais E, Teyton L, Wilson IA. The crystal structure of CD8 in complex with YTS156.7.7 Fab and interaction with other CD8 antibodies define the binding mode of CD8 alphabeta to MHC class I.. J Mol Biol. 2008; 384(5):1190-202. (Clone-specific: Blocking). View Reference
  5. Wei Y, Chen K, Sharp GC, Yagita H, Braley-Mullen H. Expression and regulation of Fas and Fas ligand on thyrocytes and infiltrating cells during induction and resolution of granulomatous experimental autoimmune thyroiditis.. J Immunol. 2001; 167(11):6678-86. (Clone-specific: In vivo exacerbation). View Reference
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569871 Rev. 1

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