16-0288-81

antibody from Invitrogen Antibodies

Targeting: CD28

Provider product page for 16-0288-81

Other assay

Antibody data

Antibody Data
Antigen structure
References [23]
Comments [0]
Validations
Other assay [2]

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Product number: 16-0288-81 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD28 Monoclonal Antibody (CD28.6), Functional Grade, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The CD28.6 monoclonal antibody reacts with the human CD28 molecule, a 44 kDa homodimer expressed by thymocytes, mature T cells and plasma cells. CD28 is a ligand for CD80 (B7-1) and CD86 (B7-2) and is a potent costimulator of T cells. Signaling through CD28 augments IL-2 and IL-2 receptor expression as well as cytotoxicity of CD3-activated T cells. The CD28.6 antibody has been reported for use in immunoblotting (WB) and neutralization studies and costimulation in the production of IL-2.
Antibody clone number: CD28.6
Concentration: 1 mg/mL

CD28 protein structure - 16-0288-81 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 LZ-8 stimulates Treg expansion in murine and human CD4 + T cells. Primary murine and human CD4 + T cells were cultured for 72 h, either in the absence of stimulation as the control (CTR), in the presence of stimulation with recombinant LZ-8 antibody (rLZ-8; 1 mu g/mL), or in the presence of stimulation with anti-CD3/28 antibodies (CD3/CD28; 1 mu g/mL each, (a, b). The cells were then harvested for flow cytometry analysis of CD25, FOXP3, and CTLA-4 expression. (c and d) The levels of IL-2, IL-10, IFN- gamma , and TGF- beta in the culture supernatants were measured by ELISA. (e) The cell proliferation of CD4 + T cells treated with or without PHA (2 mu g/mL) and then cocultured with indicated CD4 + T cells was monitored by cell-trace CFSE fluorescence via flow cytometry.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 mTOR inhibition impairs human T cell differentiation. a Titration of mTORC1 inhibitor RAD001 and dual mTORC1/2 inhibitor PP242 in human CD4 + T cells. Human naive CD4 + T cells were isolated from PBMCs of healthy donors as shown in Supplementary Fig. 1a . Isolated CD4 + T cells were treated with plate-bound alphaCD3 antibody, soluble alphaCD28, and 150 U/ml IL-2, treated with increasing doses of the indicated drugs or DMSO, for 4 d and equal amounts of whole-cell lysates analyzed by immunoblot. A representative immunoblot from one donor is shown. Four independent donors were evaluated. b RAD001 mTORC1 inhibition +TGF-beta treatment (TGFbeta in figures) increases CD4 + FOXP3 + T cells. Isolated CD4 + T cells were activated and expanded for 13 d in the presence of DMSO, 1 muM PP242, 20 nM RAD001, 2 ng/ml TGF-beta or TGF-beta plus mTOR inhibitors with 150 U/ml IL-2 as needed. At 6 and 13 d post activation, cells were stained for CD4, CD25, FOXP3, and viability analyzed by flow cytometry. P values were determined by statistical analysis using one-way ANOVA tests with Dunnett post-ANOVA test determination. Values shown are mean with standard error of the mean (SEM) of three independent studies. c mTORC2 is required for T-cell proliferation. CD4 + T cells were activated, treated, and expanded as described in b . Shown is a time course of total cell number (cell concentration x culture volume). Data represent three independent studies with SEM shown. Source data are provided