Human Survivin ELISA Kit

Catalog No.: BEK1199

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 2 pg/ml

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

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

Introduction

Survivin, also called baculoviral inhibitor of apoptosis repeat-containing 5 or BIRC5, is an inhibitor of apoptosis (IAP), or programmed cell death, that is selectively overexpressed in common human cancers. It is the smallest member of the inhibitor of apoptosis (IAP) family. It is expressed highly in most human tumours and fetal tissue, but is completely absent in terminally differentiated cells. The survivin can inhibit caspase activation, thereby leading to negative regulation of apoptosis or programmed cell death. The overexpression of survivin in tumors may serve to increase the tumors resistance to apoptosis and, thus, contribute to cell immortality even in the presence of death stimuli.

Principle of the Assay  

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

Kit components

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