Human ErbB-2 ELISA Kit

Catalog No.: BEK1052

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 10 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months.

Application: For quantitative detection of ErbB-2 in human serum, plasma, body fluids tissue lysates or cell culture supernates.

Introduction

ErbB-2 also known as HER2 (Human Epidermal Growth Factor Receptor 2), Neu, CD340 or p185, is a 185-kD glycoprotein with tyrosine kinase activity, and it is a member of the epidermal growth factor receptor (EGFR/ErbB) family. ErbB-2 is encoded by ERBB2, a known proto-oncogene located at the long arm of human chromosome 17(17q21-q22). ErbB-2 was named for its similarity to ERBB (avian erythroblastosis oncogene B). The ErbB family is composed of four plasma membrane-bound receptor tyrosine kinases. Unlike the other family members, ErbB-2 is considered to be an orphan receptor as it has no known ligand. Overexpression of ERBB2 and ERBB3 has been implicated in the neoplastic transformation of prostate cancer.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-ErbB-2 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-ErbB-2 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were 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 ErbB-2 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ErbB-2 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human ErbB-2 antibody
2. Lyophilized Human ErbB-2 standards: 2 tubes (10ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human ErbB-2 antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L