Clone:
REA612
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
T44, Tp44

Extended validation for CD28 Antibody, anti-human, 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 clonesOverlap in epitope recognition with REA612
CD28.2++
L293++
10F3n/a
15E8++
REAL105++
Cells were incubated with an excess of purified unconjugated CD28 (REA612) 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 CD28. Human peripheral blood mononuclear cells (PBMCs) 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.
View details
Flow cytometric comparison of different clones for CD28. Human peripheral blood mononuclear cells (PBMCs) 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.
View details
Flow cytometric comparison of different clones for CD28. Human peripheral blood mononuclear cells (PBMCs) 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.
View details
Flow cytometric comparison of different clones for CD28. Human peripheral blood mononuclear cells (PBMCs) 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.
View details
Flow cytometric comparison of different clones for CD28. Human peripheral blood mononuclear cells (PBMCs) 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.
View details
Flow cytometric comparison of different clones for CD28. Human peripheral blood mononuclear cells (PBMCs) 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. Human peripheral blood mononuclear cells (PBMCs) 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD28 (REA612). 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 CD28 (REA612). 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 (REA612). 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-human, REAfinity™

Overview

Clone REA612 recognizes the human CD28 antigen, a type I transmembrane protein, which is highly expressed on CD3
+
thymocytes, most peripheral thymocytes, and plasma cells but not in less mature B cells. Binding of CD28 to its ligands CD80 or CD86 costimulates T cell effector function and T cell–dependent antibody production
in vitro
and
in vivo
. In the presence of antibodies directed against CD2 and CD3, CD28 antibodies stimulate T cell proliferation and cytokine production.
Additional information: Clone REA612 displays negligible binding to Fc receptors.

Alternative names

T44, Tp44

Detailed product information

Technical specifications

CloneREA612
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD28
Alternative names of antigenT44, Tp44
Molecular mass of antigen [kDa]23
Distribution of antigenB cells, lymphocytes, T cells, plasma cells, thymocytes
Entrez Gene ID940
RRIDAB_2751558, AB_2733368, AB_2733369, AB_2733110, AB_2733111, AB_2784108, AB_2784107, AB_2802065, AB_2802058, AB_2811384, AB_2877015, AB_2877016, AB_2751569, AB_2656961, AB_2656962, AB_2904911, AB_2904910, AB_2904909, AB_2904908

Resources for CD28 Antibody, anti-human, REAfinity™

Certificates

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

References for CD28 Antibody, anti-human, REAfinity™

Publications

  1. Kozbor D. J. et al. (1987) Tp44 molecules involved in antigen-independent T cell activation are expressed on human plasma cells. J. Immunol. 138(12): 4128-4132
  2. Bahlis, N. J. et al. (2007) CD28-mediated regulation of multiple myeloma cell proliferation and survival. Blood 109(11): 5002-5010
  3. Krummel, M. F. and Allison, J. P. (1995) CD28 and CTLA-4 have opposing effects on the response of T cells to stimulation. J. Exp. Med. 182(2): 459-465
  4. Freeman, G. J. et al. (1993) Cloning of B7-2: a CTLA-4 counter-receptor that costimulates human T cell proliferation. Science 262(5135): 909-911
  5. Linsley, P. S. et al. (1993) The role of the CD28 receptor during T cell responses to antigen. Annu. Rev. Immunol. 11: 191-212
  6. June, C. H. et al. (1990) Role of the CD28 receptor in T-cell activation. Immunol. Today 11(6): 211-216

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