Human MICA ELISA Kit

Catalog No.: BEK1150

Size: 96T

Range: 62.5 pg/ml-4000 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 MICA in tissue lysates or cell culture supernates, human serum and plasma should be validated by end

users.

Introduction

MHC class I polypeptide-related sequence A(MICA) is a protein that in humans is encoded by the MICA gene. The MICA gene is located near HLA-B on chromosome 6. Nalabolu et al. (1996) showed that the MICA and MICB (602436) genes occur in a 200-kb region spanning the TNFA (191160) and TNFB (153440) cluster at chromosome 6p21.3. The protein product is expressed on the cell surface, although unlike canonical class I molecules does not seem to associate with beta-2-microglobulin. It is thought that MICA functions as a stress-induced antigen that is broadly recognized by NK cells, NKT cells, and most of the subtypes of T cells. MICA is the ligand for NK cell activating receptor NKG2D.

Principle of the Assay  

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

Kit components

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