Human CA125 / MUC16 ELISA Kit

Catalog No.: BEK1491

Size: 96T

Range: 15.6 pg/ml - 1000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of CA125 / MUC16 in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Mucin-16 (MUC-16) also known as Ovarian cancer-related tumor marker CA125 is a protein that in humans is encoded by the MUC16 gene. MUC-16 is a member of the mucin family glycoproteins. MUC-16 has found application as a tumor marker or biomarker that may be elevated in the blood of some patients with specific types of cancers, most notably ovarian cancer, or other conditions that are benign. Mucin 16 is a membrane associated mucin that possesses a single transmembrane domain. A unique property of MUC16 is its large size. MUC16 is more than twice as long as MUC1 and MUC4 and contains about 22,000 amino acids, making it the largest membrane-associated mucin.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- CA125 / MUC16 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- CA125 / MUC16 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 CA125 / MUC16 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of CA125 / MUC16 can be calculated.

Kit components

1.       One 96-well plate pre-coated with anti-human CA125 / MUC16 antibody

2.       Lyophilized human CA125 / MUC16 standards: 2 tubes (10 ng / tube)

3.       Sample / Standard diluent buffer: 30ml

4.       Biotin conjugated anti-human CA125 / MUC16 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