Clone:
H35-17.2
Type of antibody:
Primary antibodies
Isotype:
rat IgG2bκ
Applications:
FC, MICS, IF, IHC
Alternative names:
Cd8b1, Ly-3, Ly-C, Lyt-3

Extended validation for CD8b Antibody, anti-mouse

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 H35-17.2
REA793++
YTS156.7.7+
CT-CD8b++
REAL405++
Cells were incubated with an excess of purified unconjugated CD8b (H35-17.2) 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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. Splenocytes from BALB/c mice were stained with CD8b antibodies and with a suitable counterstaining. As a control, CD8b 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 (H35-17.2). 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 (H35-17.2). 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 (H35-17.2). 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 Antibody, anti-mouse

Overview

Clone H35-17.2 recognizes both alloantigeneic forms of the mouse CD8b antigen, a single-pass type I membrane protein also known as lymphocyte antigen 3 (Ly-3 or Lyt-3). The CD8 molecule is composed of two distinct polypeptide chains that pair on the cell surface either as a CD8aa homodimer or as a CD8ab heterodimer. These forms of the CD8 molecule are differentially expressed on functionally distinct CD8
+
lymphocyte subsets. The CD8ab heterodimer is expressed on cytotoxic T cells and thymocytes.

Alternative names

Cd8b1, Ly-3, Ly-C, Lyt-3

Detailed product information

Technical specifications

CloneH35-17.2
Clonalitymonoclonal
Isotyperat IgG2bκ
Isotype controlIsotype Control Antibody, rat IgG2b
Hostrat
Type of antibodyPrimary antibodies
Speciesmouse
AntigenCD8b
Alternative names of antigenCd8b1, Ly-3, Ly-C, Lyt-3
Molecular mass of antigen [kDa]22
Distribution of antigenT cells, lymphocytes, thymocytes
Entrez Gene ID12526
RRIDAB_2876918, AB_2659553, AB_2659554, AB_2659557, AB_2659558, AB_2659559, AB_2659560, AB_2659561, AB_2659562, AB_2659565, AB_2659566, AB_2659567, AB_2659568, AB_2659569, AB_2659570, AB_2811665

Resources for CD8b Antibody, anti-mouse

Certificates

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

References for CD8b Antibody, anti-mouse

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. Blanc, D. et al. (1998) Gene transfer of the Ly-3 chain gene of the mouse CD8 molecular complex: co-transfer with the Ly-2 polypeptide gene results in detectable cell surface expression of the Ly-3 antigenic determinants. Eur. J. Immunol. 18(4): 613-619

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