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Human Mast Cell Chymase ELISA Kit

Product Details

Description

CUSABIO’s Human Mast Cell Chymase ELISA Kit is a sandwich immunoassay specifically designed and validated for the quantitative detection of mast cell Chymase in the serum, cell culture supernates, or tissue homogenates. It is not intended for diagnostic use. This assay kit was designed and optimized for cardiovascular research use in humans. The kit has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details.

This assay is based on the Sandwich-ELISA mechanism (the mast cell Chymase in the sample is sandwiched between the pre-coated mast cell Chymase antibody and Biotin-labeled mast cell Chymase antibody) and enzyme-substrate chromogenic reaction. The color intensity is positively related to the amount of mast cell Chymase bound in the initial step. The levels of mast cell Chymase in the samples can be determined by referring to the O.D. (optical density) of the samples to the standard curve.

Mast cell chymase is a protease produced and released by activated mast cells. It is involved in multiple biological processes, including the conversion of angiotensin-I (Ang-I) into angiotensin-II (Ang-II), production of fibronectin and TGF-β from the extracellular matrix (ECM), processing of collagen for fibril formation from type I procollagen, and activation of matrix metalloproteinases (MMPs) from their zymogens and IL-1β, IL-18, endothelin-1, and endothelin-2 from their latent forms. Mast cell chymase also breaks down lipoproteins, facilitating the formation of macrophage foam cells. It also participates in collagen synthesis and tissue fibrosis, angiogenesis, and immunoglobulin molecule synthesis

Target Name chymase 1, mast cell
Alternative Names Alpha-chymase ELISA Kit; Chymase 1 ELISA Kit; Chymase 1 mast cell ELISA Kit; chymase 1 preproprotein transcript E ELISA Kit; chymase 1 preproprotein transcript I ELISA Kit; Chymase ELISA Kit; Chymase; heart ELISA Kit; Chymase; mast cell ELISA Kit; CMA1 ELISA Kit; CMA1_HUMAN ELISA Kit; CYH ELISA Kit; CYM ELISA Kit; EC 3.4.21.39 ELISA Kit; Mast cell chymase 1 ELISA Kit; Mast cell protease 3 ELISA Kit; Mast cell protease 5 ELISA Kit; Mast cell protease I ELISA Kit; Mast cell protease III ELISA Kit; Mcp-5 ELISA Kit; MCP3P ELISA Kit; Mcpt5 ELISA Kit; MCT1 ELISA Kit; MGC119890 ELISA Kit; MGC119891 ELISA Kit; MMCP-5 ELISA Kit
Abbreviation CMA1
Species Homo sapiens (Human)
Sample Types serum, cell culture supernates, tissue homogenates
Detection Range 0.156 ng/mL-10 ng/mL
Sensitivity 0.039 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cardiovascular
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%   
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%   
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human mast cell chymase (CMA1) in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 98  
Range % 94-103  
1:2 Average % 90  
Range % 84-95  
1:4 Average % 99  
Range % 94-104  
1:8 Average % 89  
Range % 85-94  
Recovery
The recovery of human mast cell chymase(CMA1) spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 90 85-95  
EDTA plasma (n=4) 101 95-107  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
10 2.735 2.789 2.762 2.656  
5 2.397 2.385 2.391 2.285  
2.5 1.832 1.869 1.851 1.745  
1.25 1.256 1.241 1.249 1.143  
0.625 0.725 0.756 0.741 0.635  
0.312 0.453 0.468 0.461 0.355  
0.156 0.289 0.281 0.285 0.179  
0 0.105 0.107 0.106    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human mast cell Chymase antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled mast cell Chymase antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibodyDiluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days