CD28 Antibody, anti-rat, REAfinity™

Name: CD28 Antibody, anti-rat, REAfinity™
Availability: InStock

CD28 Antibody, anti-rat, REAfinity™Extended ValidationRecombinant

Clone:

REA206

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

CD28RNA

Flow cytometry applications forCD28 Antibody, anti-rat, REAfinity™

APC
Biotin
FITC
PE
PE-Vio 770

Conjugate:

APC

Recommended dilution:

1:10

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-103-331, 130-102-692

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Lewis rat splenocytes were stained with CD28 antibodies 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 fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Extended validation for CD28 Antibody, anti-rat, 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 clones	Overlap in epitope recognition with REA206
JJ319	++
Cells were incubated with an excess of purified unconjugated CD28 (REA206) 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 CD28. Wistar rat spelnocytes were stained with CD28 antibodies and with a suitable counterstaining. As a control, CD28 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 CD28. Wistar rat spelnocytes were stained with CD28 antibodies and with a suitable counterstaining. As a control, CD28 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 CD28. Wistar rat spelnocytes were stained with CD28 antibodies and with a suitable counterstaining. As a control, CD28 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 CD28 (REA206). 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 CD28 (REA206). 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 CD28 Antibody, anti-rat, REAfinity™

Overview

Clone REA206 recognizes rat CD28, a 44 kDa glycoprotein also known as Tp44. CD28 is a homodimeric type I transmembrane protein of the Ig receptor superfamily, composed of disulfide-linked subunits. It is a costimulatory molecule which is expressed on most thymocytes, CD4

⁺

and CD8

⁺

T cells, and NK cells. Expression of CD28 is down-regulated at late stages of terminal effector T cell differentiation. Ligation of CD28 with CD80 (B7-1) and CD86 (B7-2) provides a costimulatory signal for T cell activation.

Additional information: Clone REA206 displays negligible binding to Fc receptors.

Alternative names

CD28RNA

Detailed product information

Technical specifications

Clone	REA206
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	rat
Antigen	CD28
Alternative names of antigen	CD28RNA
Molecular mass of antigen [kDa]	23
Distribution of antigen	T cells
Entrez Gene ID	25660
RRID	AB_2656942, AB_2656943, AB_2656944, AB_2656945, AB_2656946, AB_2656947, AB_2656948, AB_2656949, AB_2656950, AB_2656941

Resources for CD28 Antibody, anti-rat, REAfinity™

Certificates

Please follow this

link

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

References for CD28 Antibody, anti-rat, REAfinity™

Publications

Clark, G. J. et al. (1992) Identification of a cDNA encoding the rat CD28 homologue. Immunogenetics 35(1): 54-57
Guo, L. et al. (2003) Simultaneous blockade of co-stimulatory signals, CD28 and ICOS, induced a stable tolerance in rat heart transplantation. Transpl Immunol. 12(1): 41-48

Applications

Extended validation