CD8b Antibody, anti-human,
REAlease
^®

Name: CD8b Antibody, anti-human, REAlease®
Availability: InStock

CD8b Antibody, anti-human,
REAlease
^®
Extended ValidationRecombinant

Clone:

REAL398

Type of antibody:

Releasable antibodies, Primary antibodies, Recombinant antibodies

Applications:

FC, MICS, IHC, IF

Alternative names:

CD8 beta chain, CD8 b chain, CD8b

Flow cytometry applications forCD8b Antibody, anti-human,
REAlease
^®

APC
FITC
PE
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Tested:

QC tested

Products used:

130-120-964, 130-120-877

Protocols:

Cell surface staining protocol using REAlease™ antibodies

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with REAlease CD8b Complexes or with the corresponding REAL Control (left images) as well as with CD3 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide (PI) fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD8b Antibody, anti-human,
REAlease
^®

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

QC tested

Species tested:

human

Products used:

130-120-964, 130-120-877

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD8b Antibody, anti-human,
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 clones	Overlap in epitope recognition with REAL398
2ST8.5H7	++
SIDI8BEE	+
REA715	++
Cells were incubated with an excess of purified unconjugated CD8b (REAL398) 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.

Flow cytometric comparison of different clones for CD8b. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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

Comparison of staining pattern on non-fixed and fixed cells using CD8b (REAL398). 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 (REAL398). 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-human,
REAlease
^®

Overview

Clone REAL398 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 REAL398 recognizes the human CD8b antigen, a single-pass type I membrane protein and member of the immunoglobulin superfamily, also known as T cell surface glycoprotein Lyt-3. The CD8 glycoprotein is expressed by thymocytes, mature T cells, and natural killer (NK) cells and has been implicated in the recognition of monomorphic determinants on major histocompatibility complex class I antigens, and in signal transduction during the course of T cell activation. Both human and rodent CD8 antigens are comprised of two distinct polypeptide chains, alpha and beta. Most of peripheral blood CD8

⁺

T lymphocytes express the CD8a/CD8b heterodimer, while CD8

⁺

CD16

⁺

natural killer cells and CD8

⁺

T cell receptor γ/δ

⁺

T lymphocytes express only the CD8a/CD8a homodimer. CD8b can therefore be used to selectively bind to CD8

⁺

T cells while excluding CD8

⁺

NK cells.

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

CD8 beta chain, CD8 b chain, CD8b

Detailed product information

Technical specifications

Clone	REAL398
Clonality	monoclonal
Isotype control	Control Antibody
Host	cell line
Type of antibody	Releasable antibodies, Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD8b
Alternative names of antigen	CD8 beta chain, CD8 b chain, CD8b
Distribution of antigen	T cells, NK cells, thymocytes, CD8+ T cells
RRID	AB_2751748, AB_2751809, AB_2751751, AB_2752211, AB_2752209, AB_2811354, AB_2811352, AB_2751808

Resources for CD8b Antibody, anti-human,
REAlease
^®

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

Applications

Extended validation