Human C2C ELISA KIT

Starting at: $696.00

  • Model: ELA-E2183h
  • 20 Units in Stock
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Human C2C ELISA KIT

Packing   96Tests                                                                     Manual

FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ ENTIRE PROCEDURE!
 

Gene Name  C2C

Protein Name  Collagen Type II Cleavage

Alternative Name

Human C2C ELISA KIT ,Human COL2 ELISA KIT ,Human Collagen Type II Cleavage ELISA KIT ,Human type II collagen ELISA KIT ,Human ANFH ELISA KIT ,Human AOM ELISA KIT ,Human COL11A3 ELISA KIT ,Human SEDC ELISA KIT ,Human STL1 ELISA KIT ,Human collagen alpha-1(II) chain ELISA KIT ,Human alpha-1 type II collagen ELISA KIT ,Human arthroophthalmopathy progressive (Stickler syndrome) ELISA KIT ,Human cartilage collagen ELISA KIT ,Human chondrocalcin ELISA KIT ,Human collagen II alpha-1 polypeptide ELISA KIT ,Human collagen type II alpha 1 ELISA KIT ,Human collagen type II alpha 1 chain ELISA KIT

 

Intended use

Human C2C ELISA KIT allows for the in vitro quantitative determination of C2C concentrations in serum, plasma, tissue homogenates, cell culture supernates or other biological fluids.

 

Test principle      

The microtiter plate provided in Human C2C ELISA KIT has been pre-coated with an C2C antibody specific to C2C. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for C2C and then avidin conjugated to Horseradish Peroxidase (HRP) is added  to  each  microplate  well  and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain C2C, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of C2C in the samples is then determined by comparing the O.D. of the samples to the standard curve.

 

REAGENTS PROVIDED

All reagents provided are stored at 2-8°C. Refer to the expiration date on the label.

 

Name

96 determinations

48 determinations

MICROTITER PLATE

12*8strips

12*4strips

STANDARD6 vial

0.3ml/vial

0.3ml/vial

SAMPLE DILUENT

6.0ml

3.0ml

ENZYME CONJUGATE

10.0ml

5.0ml

WASH SOLUTION

25ml

15ml

SUBSTRATE A

6.0ml

3.0ml

SUBSTRATE B

6.0ml

3.0ml

STOP SOLUTION

6.0ml

3.0ml

Closure plate membrane

2

2

User manual

1

1

Sealed bags

1

1

Note:

1.  Standard concentration was followed by: 200, 100, 50, 25, 12.5, 6.25 ng/ml.
2.  If samples generate values higher than the highest standard, please dilute the samples with Sample Diluent and repeat the assay.

ASSAY PROCEDURE

1.  Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microtiter plate.

2.  Add 50μl of Standard or Sample to the appropriate wells. Blank well doesn’t add anyting.

3.  Add 10l of Enzymeconjugate to standard wells and sample wells except the blank well, cover with an adhesive strip and incubate for 60 minutes at 37°C.

4.  Wash the Microtiter Plate 4 times.

Manual Washing - Remove incubation mixture by aspirating contents of the plate into a sink or proper waste container. Using a squirt bottle, fill each well completely with Wash Solution (1X), then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure for a total of four times. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame.

Automated Washing - Aspirate all wells, then wash plates four times using Wash Buffer (1X). Always adjust your washer to aspirate as much liquid as possible and set fill volume at 350μL/well/wash. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears.

5.  Add Substrate A 50μl and Substrate B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.

6.  Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.

7.  Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.

 

CALCULATION OF RESULTS

  1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (X) axis versus the corresponding concentration on the horizontal (Y) axis.
  2. First, calculate the mean O.D. value for each standard and sample. All O.D. Values are subtracted by the mean value of the balnk well before result interpretation. Construct the standard curve using graph paper or statistical software.
  3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration.
  4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.
  5. Intra-assay CV(%) is less than 10% and Inter-assay CV(%) is less than 15%.
  6. Assay range: 6.25 ng/ml – 200 ng/ml.

7.  Sensitivity: The minimum detectable dose of Human C2C is typically less than 1.0 ng/ml.

8.  Cross-reactivity: This assay recognizes recombinant and natural Human C2C. No significant cross-reactivity or interference was observed.

9.  Storage: 2-8 (Use frequently); six months (-20).

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