Human P53 ELISA Kit

Catalog No.: BEK1183

Size: 96T

Range: 156 pg/ml-10,000 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 P53 in human serum, plasma, cell culture supernatants or cell lysate.

Introduction

P53, also known as protein 53 or tumor protein 53, named in reference to its apparent molecular mass: It runs as a 53-kilodalton (kDa) protein on SDS-PAGE. Human p53 is 393 amino acids long and has seven domains. It plays a role in conserving stability by preventing genome mutation, as such, p53 has been described as "the guardian of the genome". p53 also has many mechanisms of anticancer function, and plays a role in apoptosis, genomic stability, and inhibition of angiogenesis.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-P53 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-P53 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 P53 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of P53 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human P53 antibody
2. Lyophilized human P53 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human P53 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 1