CD28 Antibody, anti-mouse, REAfinity™

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

CD28 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA806

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

T44, Tp44

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

APC
Biotin
PE

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-111-973, 130-111-820

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Splenocytes from BALB/c mice were stained with CD28 antibodies or with the corresponding REA Control antibodies (left image) as well as with CD3ε antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Extended validation for CD28 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 clones	Overlap in epitope recognition with REA806
E18	-
37.51	++
Cells were incubated with an excess of purified unconjugated CD28 (REA806) 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. C57BL/6 mouse Splenocytes 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. C57BL/6 mouse Splenocytes 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. C57BL/6 mouse Splenocytes 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. C57BL/6 mouse Splenocytes 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 (REA806). 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 (REA806). 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-mouse, REAfinity™

Overview

Clone REA806 recognizes the mouse CD28 antigen, a costimulatory molecule, expressed on most thymocytes, CD4

⁺

and CD8

⁺

T cells, and NK cells. Expression is down-regulated at late stages of terminal effector T cell differentiation. CD28 is a homodimeric type I transmembrane protein of the Ig receptor superfamily, composed of disulfide-linked 45 kDa subunits. Ligation of CD28 with CD80 (B7-1) and CD86 (B7-2) provides a costimulatory signal for T cell activation.

Additional information: Clone REA806 displays negligible binding to Fc receptors.

Alternative names

T44, Tp44

Detailed product information

Technical specifications

Clone	REA806
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	mouse
Antigen	CD28
Alternative names of antigen	T44, Tp44
Molecular mass of antigen [kDa]	23
Distribution of antigen	T cells, B cells, lymphocytes, thymocytes
Entrez Gene ID	12487
RRID	AB_2656964, AB_2656965, AB_2656966, AB_2656967, AB_2656963

Resources for CD28 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this

link

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

References for CD28 Antibody, anti-mouse, REAfinity™

Publications

Gross, J. A. et al. (1990) The murine homologue of the T lymphocyte antigen CD28. Molecular cloning and cell surface expression. J Immunol 144(8): 3201-3210
Harper, K. et al. (1991) CTLA-4 and CD28 activated lymphocyte molecules are closely related in both mouse and human as to sequence, message expression, gene structure, and chromosomal location. J Immunol 147(3): 1037-1044

Applications

Extended validation