CD28 ELISA Kit

Catalog No. ABIN5510635

This Colorimetric ELISA kit is designed for the quantitative measurement of Human CD28.

Quick Overview for CD28 ELISA Kit (ABIN5510635)

Target: CD28

Binding Specificity: AA 19-152

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5 pg/mL - 4000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma (EDTA), Plasma (heparin), Serum

Product Details CD28 ELISA Kit

Minimum Detection Limit: 62.5 pg/mL

Purpose: Sandwich Human CD28 ELISA Kit to quantitate Human CD28 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Analytical Method: Quantitative

Specificity: Expression system for standard: NS0
Immunogen sequence: N19-P152

Capture antibody and Detection antibody: a monoclonal antibody from mouse, a biotinylated polyclonal antibody from goat

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: < 12 pg/mL

Components:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Material not included: Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional). Pipettes and pipette tips capable of precisely dispensing 0.5 μL through 1 mL volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500 mL graduated cylinders. Test tubes for dilution.

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Sample Volume: 100 μL

Plate: Pre-coated

Protocol: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the target amount in sample captured in plate.

Assay Precision: Intra-Assay Precision (CV%): 7.1%, 4.5%, 6%
Inter-Assay Precision (CV%): 8.6%, 5.6%, 6.1%

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Expiry Date: 12 months

Target Details for CD28

Target: CD28

Alternative Name: CD28

Background:

Background: 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). The CD28 gene is mapped to 2q33-q34. CD28 and CTLA-4, together with their ligands, B7-1 and B7-2, constitute one of the dominant costimulatory pathways that regulate T and B cell responses. CD28 and CTLA-4 are structurally homologous molecules that are members of the immunoglobulin (Ig) gene superfamily.

Gene Full Name: CD28 Molecule

Gene ID: 940

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