Recombinant CD28 antibody

Catalog No. ABIN7543356

The Rabbit Chimeric anti-CD28 antibody has been validated for IF, IP, ELISA and BP. It is suitable to detect CD28 in samples from Human.

Quick Overview for Recombinant CD28 antibody (ABIN7543356)

Target: CD28

Antibody Type: Recombinant Antibody

Reactivity: Human

Host: Rabbit

Clonality: Chimeric

Conjugate: This CD28 antibody is un-conjugated

Application: Immunofluorescence (IF), Immunoprecipitation (IP), ELISA, Blocking Peptide (BP)

Clone: 9-3

Product Details anti-CD28 Antibody

Purpose: Anti-CD28 [9.3], Rabbit IgG, kappa

Specificity: The antibody is specific for human CD28. CD28 is one of the proteins expressed on T cells that provide co-stimulatory signals required for T cell activation and survival. T cell stimulation through CD28 in addition to the T-cell receptor (TCR) can provide a potent signal for the production of various interleukins.

Characteristics:

Original Species of Ab: Mouse

Original Format of Ab: IgG2a

Purification: Protein A affinity purified

Immunogen: The original antibody was generated by immunizing mice with human T cells.

Isotype: IgG kappa

Application Details

Application Notes: The antibody was found to induce T cell activation and proliferation in collaboration with TPA. Further, T cell proliferation was associated with IL-2 receptor expression and IL-2 production. The antibody stained 60-80 % of the isolated T cells by immunofluorescence. The antibody immunoprecipitated CD28 from Jurkat cells. (Hara et al., 1985, PMID: 3159820). Immunofluorescence was performed on T cells expressing CD28 using this antibody. The antibody was cytotoxic for 50-80 % of peripheral T cells and for 20-50 % of thymocytes but it was not cytotoxic for cultured B-cell lines, normal B cells, or monocytes. In immune precipitation assays, the antibody reacted with CD28 (Hansen et al., 1980). The antibody induces proliferative responses in human T cells in in vitro experiments (Hutloff et al. 1999, PMID: 9930702). The antibody detected CD28 by western blot analysis (Siefken et al,1998, PMID: 9712026). Humanize, chimeric and hybrid Fab versions of the antibody were constructed. The ability of the three Fabs to bind to CD28 was examined by ELISA, the three Fabs showed a binding activity, with an EC50 of 630, 20 and 30 nM respectively for the humanized, the chimeric, hybrid Fab version. Thus, the humanized Fab showed a moderate loss in avidity. The three Fab versions of the antibody were tested for blocking ligation of CD28-Ig to CHO cells expressing CD80 and the binding was quantified by flow cytometry. The chimeric Fab exhibited the best blocking activity followed by the hybrid and the humanized Fab, with EC50 values of 0.2, 0.4, and 1.2 μM, respectively. Lastly, three Fab versions showed biological activity as an immunosuppressant by inhibiting a MLR (Tan et al. 2002, PMID: 12097421).

Comment:

This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG2a format for improved compatibility with existing reagents assays and techniques.

Restrictions: For Research Use only

Handling

Concentration: 1 mg/mL

Buffer: PBS with 0.02 % Proclin 300.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C.

Target Details for CD28

Target: CD28

Alternative Name: CD28

Background: T-cell-specific surface glycoprotein CD28, TP44

UniProt: P10747

Pathways: TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response