Active Member
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[China]
Address: #1008.Junjiang Internatioanl Bldg. 228 Ningguo Rd. Yangpu Dist. Shanghai. 200090. China
Contact name:
Shanghai Korain Biotech Co., Ltd |
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Customized Canine MCP-1 CCL2 Strong Sensitivity MCAF ELISA Test Kit
With 2 Hours Assay Time
Assay Principle
This ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The
plate has been pre-coated with Canine MCP-1/CCL2/MCAF antibody.
MCP-1/CCL2/MCAF present in the sample is added and binds to
antibodies coated on the wells. And then biotinylated Canine
MCP-1/CCL2/MCAF Antibody is added and binds to MCP-1/CCL2/MCAF in
the sample. Then Streptavidin-HRP is added and binds to the
Biotinylated MCP-1/CCL2/MCAF antibody. After incubation unbound
Streptavidin-HRP is washed away during a washing step. Substrate
solution is then added and color develops in proportion to the
amount of Canine MCP-1/CCL2/MCAF. The reaction is terminated by
addition of acidic stop solution and absorbance is measured at 450
nm.
Intended Use
This sandwich kit is for the accurate quantitative detection of
Canine Monocyte chemotactic protein 1,monocyte chemotactic and
activating factor (also known as MCP-1/CCL2/MCAF) in serum, plasma,
cell culture supernates, cell lysates, tissue homogenates.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of
known concentration were tested on one plate to assess intra-assay
precision.
Inter-Assay Precision (Precision between assays) Three samples of
known concentration were tested in separate assays to assess
inter-assay precision.
CV(%) = SD/mean x 100
Intra-Assay: CV<8%
Inter-Assay: CV<10%
Reagent Provided
| Components | Quantity |
| Standard Solution (1280ng/L) | 0.5ml x1 |
| Pre-coated ELISA Plate | 12 * 8 well strips x1 |
| Standard Diluent | 3ml x1 |
| Streptavidin-HRP | 6ml x1 |
| Stop Solution | 6ml x1 |
| Substrate Solution A | 6ml x1 |
| Substrate Solution B | 6ml x1 |
| Wash Buffer Concentrate (25x) | 20ml x1 |
| Biotinylated Canine MCP-1/CCL2/MCAF Antibody | 1ml x1 |
| User Instruction | 1 |
| Plate Sealer | 2 pics |
| Zipper bag | 1 pic |
Precautions
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (1280ng/L) with 120μl of
standard diluent to generate a 640ng/L standard stock solution.
Allow the standard to sit for 15 mins with gentle agitation prior
to making dilutions. Prepare duplicate standard points by serially
diluting the standard stock solution (640ng/L) 1:2 with standard
diluent to produce 320ng/L, 160ng/L, 80ng/L and 40ng/L solutions.
Standard diluent serves as the zero standard(0 ng/L). Any remaining
solution should be frozen at -20°C and used within one month.
Dilution of standard solutions suggested are as follows:
| 640ng/L | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 320ng/L | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 160ng/L | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 80ng/L | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 40ng/L | Standard No.1 | 120μl Standard No.2 + 120μl Standard Diluent |
| Standard Concentration | Standard No.5 | Standard No.4 | Standard No.3 | Standard No.2 | Standard No.1 |
| 1280ng/L | 640ng/L | 320ng/L | 160ng/L | 80ng/L | 40ng/L |
Wash Buffer Dilute 20ml of Wash Buffer Concentrate 25x into deionized or
distilled water to yield 500 ml of 1x Wash Buffer. If crystals have
formed in the concentrate, mix gently until the crystals have
completely dissolved.
Summary
1. Prepare all reagents, samples and standards.
2. Add sample and ELISA reagent into each well. Incubate for 1 hour
at 37°C.
3. Wash the plate 5 times.
4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.
5. Add stop solution and color develops.
6. Read the OD value within 10 minutes.
Cat.No E0135Ca
Standard Curve Range: 5ng/L - 1000ng/L
Sensitivity: 2.55ng/L
