Human KIM-1 ELISA Kit

Catalog No.: BEK1130

Size: 96T

Range: 31.2 pg/ml-2000 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of KIM-1 in human serum, plasma, urine, cell culture supernatant or tissue samples.

Introduction

KIM1, also known as Hepatitis A virus cellular receptor 1, is a protein that in humans is encoded by the HAVCR1 gene, which maps to 5q33.2. It is a major cause of orally transmitted acute hepatitis, infects primate cells, but not dog or rat cells, after binding to the HAV cellular receptor (HAVCR). Infection of canine osteogenic sarcoma cells expressing HAVCR1 with HAV led Feigelstock et al. (1998) to conclude that the protein is indeed a receptor for the virus. Khademi et al. (2004) found that differential expression of TIMs by Th1 and Th2 cells may be implicated in different phases of an autoimmune disease.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti-KIM-1 antibody was pre-coated onto 96-well plates. And the HRP conjugated anti-KIM-1 antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 KIM-1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of KIM-1 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human KIM-1 antibody

2. Standard: 0.5ml (2700 pg /mL)

3. Standard diluent buffer:  1.5 ml

4. Wash buffer (30×):  20 ml. Dilution: 1:30

5. Sample diluent buffer: 6 ml

6. HRP conjugated anti-human KIM-1 antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A:  6ml

9. TMB substrate B:  6ml

10. Plate sealer:   2

11. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L