Clone:
REA865
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
Lyt-3.2, Ly-3.2

Extended validation for CD8b.2 Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA865
53-5.8++
Cells were incubated with an excess of purified unconjugated CD8b.2 (REA865) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD8b.2. C57BL6/6 splenocytes were stained with CD8b.2 antibodies and with a suitable counterstaining. As a control, CD8b.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD8b.2. C57BL6/6 splenocytes were stained with CD8b.2 antibodies and with a suitable counterstaining. As a control, CD8b.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD8b.2. C57BL6/6 splenocytes were stained with CD8b.2 antibodies and with a suitable counterstaining. As a control, CD8b.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD8b.2. C57BL6/6 splenocytes were stained with CD8b.2 antibodies and with a suitable counterstaining. As a control, CD8b.2 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD8b.2 (REA865). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD8b.2 (REA865). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD8b.2 (REA865). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD8b.2 Antibody, anti-mouse, REAfinity™

Overview

Clone REA865 recognizes the β chain of the mouse CD8 antigen, a single-pass type I membrane protein also known as lymphocyte antigen 3 (Ly-3.2 or Lyt-3.2). The CD8 molecule is composed of two distinct polypeptide chains that pair on the cell surface either as a CD8 αα homodimer or as a CD8 αβ heterodimer. These forms of the CD8 molecule are differentially expressed on functionally distinct CD8
+
lymphocyte subsets. The CD8 αβ heterodimer is expressed on cytotoxic T cells and thymocytes. CD8b functions as a co-receptor for the T cell receptor (TCR).
Additional information: Clone REA865 displays negligible binding to Fc receptors.

Alternative names

Lyt-3.2, Ly-3.2

Detailed product information

Technical specifications

CloneREA865
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD8b.2
Alternative names of antigenLyt-3.2, Ly-3.2
Molecular mass of antigen [kDa]22
Distribution of antigenT cells, thymocytes
Entrez Gene ID12526
RRIDAB_2726530, AB_2726600, AB_2726531, AB_2726601, AB_2726532, AB_2726605, AB_2726536, AB_2726602, AB_2726533, AB_2726603, AB_2726534, AB_2726604, AB_2726535, AB_2726598, AB_2726529, AB_2726599

Resources for CD8b.2 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD8b.2 Antibody, anti-mouse, REAfinity™

Publications

  1. Chang, H. C. et al. (2005) Structural and mutational analyses of a CD8alphabeta heterodimer and comparison with the CD8alphaalpha homodimer. Immunity 23(6): 661-671
  2. Renard, V. et al. (1996) CD8 beta increases CD8 coreceptor function and participation in TCR-ligand binding. J. Exp. Med. 184(6): 2439-2444
  3. Gao, G. F. et al. (2000) Molecular interactions of coreceptor CD8 and MHC class I: the molecular basis for functional coordination with the T-cell receptor. Immunol. Today 21(12): 630-636

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