Clone:
REAL405
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC
Alternative names:
Cd8b1, Ly-3, Ly-C, Lyt-3

Extended validation for CD8b Antibody, anti-mouse,
REAlease
®

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 REAL405
H35-17.2++
YTS156.7.7+
CT-CD8b++
REA793++
Cells were incubated with an excess of purified unconjugated CD8b (REAL405) 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 (REAL405). 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 (REAL405). 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 (REAL405). 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,
REAlease
®

Overview

Clone REAL405 is an antibody fragment derived from the full CD8b antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.
Clone REAL405 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.
The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Alternative names

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

Detailed product information

Technical specifications

CloneREAL405
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD8b
Alternative names of antigenCd8b1, Ly-3, Ly-C, Lyt-3
Distribution of antigenT cells, thymocytes, CD8+ T cells
RRIDAB_2801729, AB_2801962, AB_2811709, AB_2811710, AB_2811708, AB_2801728

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