Human Tryptase ELISA Kit

Catalog No.: BZEK2356

Size: 96T

Range: 1.5 ng/mL - 24 ng/mL

Sensitivity: 0.1 ng/mL

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

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

Introduction

Mast cells are involved in a number of diseases, including inflammatory diseases such as asthma. Tryptase is a known marker of mast cell burden and activity. Sverrild A et al conclude that as much as 82% of the variation in serum tryptase is due to genetic factors. Body mass index and sex, but not asthma or airway hyperresponsiveness (AHR) to methacholine, correlate to serum tryptase. A genetic overlap may exist between serum tryptase and body mass index. Tryptase(+) mast cells (MCs), abundant in the invasive front of tumours, contribute to tissue remodelling. Indeed, protease-activated receptor-2 (PAR-2) activation by MC-tryptase is considered an oncogenic event in colorectal cancer (CRC). The high density of tryptase(+) MCs at invasive margins of tumours was associated with advanced stages of CRC and was strongly correlated with PAR-S expression.

Principle of the Assay  

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

Kit components

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

2. Standard: 0.5ml (48 ng/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 tryptase 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