Human COL1A1 ELISA Kit

Catalog No.: BEK1294

Size: 96T

Range: 31.2 pg/ml - 2000 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 COL1A1 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Collagen, type I, alpha 1, also known as alpha-1 type I collagen, is a protein that in humans is encoded by the COL1A1 gene. COL1A1 encodes the major component of type I collagen, the fibrillar collagen found in most connective tissues, including cartilage. The COL1A1 gene produces a component of type I collagen, called the pro-alpha1(I) chain. This chain combines with another pro-alpha1(I) chain and also with a pro-alpha2(I) chain (produced by the COL1A2 gene) to make a molecule of type I procollagen. These triple-stranded, rope-like procollagen molecules must be processed by enzymes outside the cell. Once these molecules are processed, they arrange themselves into long, thin fibrils that cross-link to one another in the spaces around cells.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- COL1A1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- COL1A1 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 COL1A1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of COL1A1 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human COL1A1 antibody
2. Lyophilized human COL1A1 standards: 2 tubes (10 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human COL1A1 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