Mouse Collagen Type Ⅰ,Col ⅠELISA Kit

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Name: Mouse Collagen Type Ⅰ,Col ⅠELISA Kit
Brand: CUSABIO
SKU: CSB-E08083m

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

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

Code	CSB-E08083m
Size	96T,5×96T,10×96T
Trial Size
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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.