Model: | 10316 |
---|---|
Brand: | Glory |
Origin: | Made In China |
Category: | Chemicals / Biochemical |
Label: | ELISA , kit , antibody |
Price: |
US $690
/ pc
|
Min. Order: | 1 pc |
Human Tissue Inhibitor of Metalloproteinase 1
(TIMP1)ELISA Kit
FORRESEARCH USE ONLY. Not for clinical diagnosis use
Catalog #:10316
Introduction
Principle of test
The kit is for the quantitative level of TIMP1 in the sample, adopt purified human TIMP1 to coat microtiter plate, make solid-phase antibody, then add samples or standards to wells with a labeled antibody specific to TIMP1, then add labeled HRP to the well. After washing completely, add TMB substrate solution, TMB substrate becomes blue color in wells that contains antibody - antigen - enzyme-antibody complex, reaction is terminated by the addition of a stop solution and the color change is measured at a wavelength of 450 nm. The concentration of TIMP1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Composition of the kit
Reagent |
Quantity |
Microelisa Stripplate |
12well×8strips |
Standard: 108μg/L |
1×0.5ml |
Standard Diluent |
1×1.5ml |
HRP-Conjugate Reagent |
1×6ml |
Sample Diluent |
1×6ml |
Chromogen Solution A |
1×6ml |
Chromogen Solution B |
1×6ml |
Stop Solution |
1×6ml |
Wash Solution |
1×20ml×30 fold |
User manual |
1 |
Adhesive Strip |
2 |
Sealed bag |
1 |
Storage conditions
Washing method
SAMPLE PREPARATION
Assay procedure
Step 1: Standard: Bring all reagents to room temperature.
Dilute the standard:Pipette 50μl standard dilution in each tube. Pipette 100μl standard (108μg/L) in the first tube. And take out 100μl from the first tube into the second. Pipette 50μl from the second tube to the third tube and produce dilution series as below. Repeat each of the concentration to get the mean value of each well.
Tube |
0 |
1 |
2 |
3 |
4 |
5 |
μg/L |
108 |
72 |
48 |
24 |
12 |
6 |
Step 2: Prepare sample: Set blank wells separately (blank comparison wells don't add sample and HRP-Conjugate reagent, other each step operation is same). Pipette Sample dilution 40μl to testing sample well, then add testing sample 10μl (sample final dilution is 5-fold), Pipette sample to wells, don’t touch the well wall as far as possible, and mix gently.
Step 3: Incubate: Cover with the adhesive strip provided, incubate for 30 min at 37℃.
Step 4: Configurate liquid: Dilute wash solution 30-fold (or 20-fold) with distilled water.
Step 5: Washing: Uncover the adhesive strip, discard liquid, Pipette washing buffer to every well, still for 30s then drain, repeat 5 times.
Step 6: Add enzyme: Pipette HRP-Conjugate reagent 50μl to each well, except blank well.
Step 7: Incubate: Operation with 3.
Step 8: Washing: Operation with 5.
Step 9: Color: Pipette Chromogen Solution A 50ul and Chromogen Solution B to each well, avoid the light preservation for 15 min at 37℃
Step 10: Stop the reaction: Pipette Stop Solution 50μl to each well, Stop the reaction (the blue change to yellow).
Step 11: Calculate: take blank well as zero, Read absorbance at 450nm after Pipetteing Stop Solution within 15min.
CALCULATION of result
Take the standard concentration as the horizontal, the OD value for the vertical ,draw the standard curve on graph paper, Find out the corresponding concentration according to the sample OD value by the Sample curve, multiplied by the dilution multiple, or calculate the straight line regression equation of the standard curve with the standard concentration and the OD value, with the sample OD value in the equation, calculate the sample concentration, multiplied by the dilution factor, the result is the sample actual concentration.
Graphical Representation as following:
DESCRIPTION
1. The standard curve is drawn under ideal conditions, and is just for reference rather than the actual standard curve diagram of the kit.
2. It is advisable to establish proper assay data and standard curve according to respective laboratory conditions.
DETECTION RANGE
Detection range of the kit is: 4μg/L -80μg/L
Expiration
Twelve months [see label on the outer box for the specific date].
Attention
Glory Science Co., Ltd. | |
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Country/Region: | Zhe Jiang - China |
Business Nature: | Trading Company |
Phone: | 86752285 |
Contact: | bellary (exporter) |
Last Online: | 15 Jan, 2015 |