NA/LE Mouse anti-human CD28 mAb

Catalog Number: S0B0010 Application: FCM,Functional/Block/Neutralize Reactivity: Human Conjugation: Unconjugated Brand: Starter

Price:

$375 USD

Size: 1mg 5mg

Quantity:

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Product Details

Product Specification

Host	Mouse
Antigen	CD28
Synonyms	TP44
Immunogen	N/A
Accession	P10747
Clone Number	15E8
Antibody Type	Mouse mAb
Isotype	IgG1,k
Isotype Control	Invivo mouse IgG1 isotype control
Application	FCM, Functional Assay, in vitro T cell stimulation/activation
Reactivity	Hu
Purification	Protein G
Concentration	Lot specific* (generally 0.5 to 10 mg/ml)*
Endotoxin	＜1EU/mg
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS pH7.4, containing no preservative
Stability & Storage	2 to 8 °C for 2 weeks under sterile conditions; -20 °C for 3 months under sterile conditions; -80 °C for 24 months under sterile conditions. Please avoid repeated freeze-thaw cycles.

Dilution

application	dilution	species
Functional Assay	1:500
FCM	1:500-1000

Background

This antibody recognizes the same epitope as clone 15E8.

CD28 (Cluster of Differentiation 28) 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 (IL-6 in particular).
CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins. When activated by Toll-like receptor ligands, the CD80 expression is upregulated in antigen-presenting cells (APCs). The CD86 expression on antigen-presenting cells is constitutive (expression is independent of environmental factors). It is generally reported, that CD28 is expressed on 50% of CD8+ T cells and more than 80% CD4+ T cells in human, but during the course of activation some T cells lose this molecule. In general, CD28 is a primary costimulatory molecule for T cell activation.

Picture

Contents & Protocol

Procedure

1. Isolate human PBMCs according to the manufacturer’s instructions.

2. Suspend PBMCs in medium at 106 cells/mL.

3. Do one of the following depending on the format you are using:

· For the soluble format, incubate 106 cells with CD3 Mouse mAb (OKT3) at 1 µg/mL final concentration in culture for 3 days (5% CO2, 37°C).

· For the immobilized format, coat the CD3 Mouse mAb (OKT3) onto a plate at 5 to 10 μg/mL at 4°C, overnight. Add cells to the CD3 Mouse mAb (OKT3)-coated plate.

4. Wash the cells twice and resuspend them in medium at 106 cells/mL.

5. Distribute the cells in a round-bottom plate at 105 cells per well.

6. [Optional for co-stimulation] Add CD28 Mouse mAb (15E8) and optionally Protein G at 5 µg/mL each final concentration, and incubate the plate for an additional 3 days (5% CO2 at 37°C).

Validation Data

Flow cytometric analysis of THP-1 (Human monocytic leukemia monocyte, left) / Jurkat (Human T cell leukemia T lymphocyte, right) cells labelling CD28 antibody at 1/500 dilution (0.1 μg)/ (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Mouse IgG Alexa Fluor® 488 was used as the secondary antibody. Negative control: THP-1

T cell proliferation Assay with CD3(S0B0009) &CD28 (S0B0010)

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NA/LE Mouse anti-human CD28 mAb

NA/LE Mouse anti-human CD28 mAb