Human ANG ELISA Kit

Catalog No.: BEK1007

Size: 96T

Range: 78 pg/ml - 5000 pg/ml

Sensitivity < 12 pg/ml

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

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

Introduction

Angiogenin (ANG) also known as ribonuclease 5, is a potent stimulator of new blood vessel formation. ANG is a member of the pancreatic ribonuclease A superfamily, and RNase activity of ANG is important for its angiogenic activity. It is expressed in the neuroaxis. Like VEGF, ANG is induced by hypoxia to elicit angiogenesis and is expressed in motor neurons. Angiogenin may function as a tRNA-specific ribonuclease that binds to actin on the surface of endothelial cells, once bound, angiogenin is endocytosed and translocated to the nucleus, thereby promoting the endothelial invasiveness necessary for blood vessel formation. Angiogenin induces vascularization of normal and malignant tissues, and abolishes protein synthesis by specifically hydrolyzing cellular tRNAs.

Principle of the Assay  

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

Kit components

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