Mouse Collagen Type Ⅰ,Col ⅠELISA Kit

Instructions
Code CSB-E08083m
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Mouse Collagen Type I (Col I) ELISA Kit is specifically designed to standardize the detection of mouse Col I in multiple biological solutions, including serum, plasma, cell culture supernates, and tissue homogenates. This kit employs the Sandwich-ELISA mechanism to quantitatively measure the concentration of Col I. The Col I in the sample and standard is bound in a "sandwich" format by the capture antibody pre-coated on the microwell plate wells and subsequently added Biotin-labeled Col I antibody. The HRP-avidin is added and conjugated to the biotinylated Col I antibody, allowing for a colorimetric reaction to ensue upon TMB substrate addition. HRP-TMB interaction yields a blue color. The color development is terminated, and solution color changes from blue to yellow. The intensity is a positive representative to the Col I concentration.

This kit has been professionally verified with reliable quality, high sensitivity, strong specificity, good linearity, recovery of 87-107%, precision less than 10%, and consistency between batches. See more details on the product instructions.

Collagen type I is the most abundant extracellular matrix protein. It is made up of two α1(I) chains and one α2(I) chain. Apart from cartilaginous tissues, collagen type I is present throughout the body, including skin, tendons, vasculature, as well as organs such as lungs, and heart. It also forms the main component in the organic portion of the calcified tissue of bone and teeth. The interaction between collagen type I and integrin α2β1 is essential for bone-resorbing cells and osteoclasts.

Target Name Collagen Type Ⅰ,Col Ⅰ
Alternative Names Collagen Type 1,Col 1,Collagen Type I,Col I
Abbreviation Col Ⅰ
Species Mus musculus (Mouse)
Sample Types serum, plasma, 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 Signal Transduction
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
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 mouse ColⅠ 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)  
1:100 Average % 92  
Range % 87-97  
1:200 Average % 96  
Range % 91-104  
1:400 Average % 97  
Range % 94-101  
1:800 Average % 98  
Range % 89-102  
Recovery
The recovery of mouse ColⅠ 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) 103 98-107  
EDTA plasma (n=4) 92 87-98  
             
             
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 2.595 2.494 2.545 2.355  
5 1.904 1.803 1.854 1.664  
2.5 1.148 1.128 1.138 0.948  
1.25 0.660 0.671 0.666 0.476  
0.625 0.432 0.442 0.437 0.247  
0.312 0.304 0.315 0.310 0.120  
0.156 0.249 0.258 0.254 0.064  
0 0.189 0.190 0.190    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-mouse Col I 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 Col I 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-antibody Diluent (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 that 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
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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