Human Creatine Kinase MB, CK-MB ELISA Kit

Catalog No.: BEK1248

Size: 96T

Range: 3.12 ng/ml-200 ng/ml

Sensitivity < 1 ng/ml

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

Application: For quantitative detection of CK-MB in human serum, plasma, tissue homogenates or cell culture supernatants.

Introduction

Creatine kinase isoenzyme-MB (CK-MB) is a biomarker for detecting myocardial injury. It is a simple, widely available, and useful biomarker for predicting adverse in-hospital clinical outcomes in pulmonary embolism. The encoded protein reversibly catalyzes the transfer of phosphate between ATP and various phosphogens such as creatine phosphate. It acts as a homodimer in striated muscle as well as in other tissues, and as a heterodimer with a similar brain isozyme in heart. The CK-MB-to-total-CK ratio is an easily available indicator and could be clinically utilized as a primary screening tool for cancer. Higher ratio of CK-MB-to-total-CK was specifically associated with certain malignancies, like colorectal cancer, lung cancer and hepatocellular carcinoma, as well as some cancer-associated status factors such as advanced stage and liver metastasis.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti-human CK-MB antibody
2. Lyophilized human CK-MB standards: 2 tubes (200 ng/tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human CK-MB 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