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Shanghai Korain Biotech Co Ltd
8 Years
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| Brand Name : | BT Lab |
| Model Number : | Cat.No E1295Mo |
| Certification : | CE, ISO9001:2005, MSDS |
| Price : | Negotiation |
| Supply Ability : | Western Union, T/T |
| Delivery Time : | 1-3 business days, bulk order within one week |
96 Wells 48 Wells COL4A1 Collagen 4Mouse Sandwich ELISA Kit With
High Sensitivity and Specificity
Cat.No E1295Mo
Standard Curve Range: 0.2ng/ml - 70ng/ml
Sensitivity: 0.116ng/ml
Size: 96 wells
*This product is for research use only, not for use in diagnosis
procedures. It’s highly recommend to read this instruction entirely
before use.
Precautions
Assay Principle
This sandwich elisa kit is an Enzyme-Linked Immunosorbent Assay
(ELISA). The plate has been pre-coated with Mouse COL4A1/Collagen 4
antibody. COL4A1/Collagen 4 present in the sample is added and
binds to antibodies coated on the wells. And then biotinylated
Mouse COL4A1/Collagen 4 Antibody is added and binds to
COL4A1/Collagen 4 in the sample. Then Streptavidin-HRP is added and
binds to the Biotinylated COL4A1/Collagen 4 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 Mouse COL4A1/Collagen 4. The reaction
is terminated by addition of acidic stop solution and absorbance is
measured at 450 nm.
Reagent Provided
| Components | Quantity |
| Standard Solution (80ng/ml) | 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 Mouse COL4A1/Collagen 4 Antibody | 1ml x1 |
| User Instruction | 1 |
| Plate Sealer | 2 pics |
| Zipper bag | 1 pic |
Material Required But Not Supplied
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (80ng/ml) with 120μl of
standard diluent to generate a 40ng/ml 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 (40ng/ml) 1:2 with standard
diluent to produce 20ng/ml, 10ng/ml, 5ng/ml and 2.5ng/ml solutions.
Standard diluent serves as the zero standard(0 ng/ml). Any
remaining solution should be frozen at -20°C and used within one
month. Dilution of standard solutions suggested are as follows:
| 40ng/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 20ng/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 10ng/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 5ng/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 2.5ng/ml | 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 |
| 80ng/ml | 40ng/ml | 20ng/ml | 10ng/ml | 5ng/ml | 2.5ng/ml |
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.
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