Human TGFα ELISA Kit

Catalog No.: BEK1200

Size: 96T

Range: 15.6 pg/ml-1000 pg/ml

Sensitivity < 1 pg/ml

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

Application: For quantitative detection of TGFα in Human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Transforming growth factor alpha (TGF-α) is upregulated in some human cancers. It is produced in macrophages, brain cells, and keratinocytes, and induces epithelial development. It is closely related to EGF, and can also bind to the EGF receptor with similar effects. TGFα stimulates neural cell proliferation in the adult injured brain. TGFA is expressed rhythmically in the suprachiasmatic nucleus, and when infused into the third ventricle it reversibly inhibited locomotor activity and disrupted circadian sleep-wake cycles. TGFA also plays a role in certain paraneoplastic manifestations of melanoma.

Principle of the Assay 

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- TGFα polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- TGFα 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 TGFα amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of TGFα can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human TGFα antibody
2. Lyophilized Human TGFα standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human TGFα 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