Human Kidney injury molecule 1, KIM-1 ELISA Kit

Catalog No.: BZEK1498-48

Size: 48T

Range: 5 ng/L - 80 ng/L

Sensitivity: 0.5 ng/L

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

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

Introduction

Hepatitis A virus cellular receptor 1 (HAVcr-1) also known as T-cell immunoglobulin and mucin domain 1 (TIM-1) is a protein that in humans is encoded by the HAVCR1 gene. It is also known as KIM-1 Kidney Injury Molecule -1, which is a protein the most highly upregulated in injured kidneys by various types of insults. Its upregulation during renal injury has been found in the kidneys of the vertebrates such as Zebrafish and humans. The hepatitis A virus cellular receptor 1 (HAVCR1/TIM-1), is a member of the TIM (T cell transmembrane, immunoglobulin, and mucin) gene family, which plays critical roles in regulating immune cell activity especially regarding the host response to viral infection. TIM-1 is also involved in allergic response, asthma, and transplant tolerance.

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 48-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 48-well plate pre-coated with anti- human KIM-1 antibody

2. Standard: 0.5ml (160 ng/L)

3. Standard diluent buffer: 1.5 ml

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

5. Sample diluent buffer: 3 ml

6. HRP conjugated anti- human KIM-1 antibody (RTU): 3ml

7. Stop solution: 3 ml

8. TMB substrate A:  3ml

9. TMB substrate B:  3ml

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