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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Rat Strong Sensitivity Thiobarbituric Acid Reactive Substance
Sandwich ELISA Kit With 2 Hours Assay Time
Cat.No E1369Ra
Standard Curve Range: 0.5nmol/ml - 100nmol/ml
Sensitivity: 0.26nmol/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.
Reagent Provided
| Components | Quantity |
| Standard Solution (128nmol/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 Rat TBARS Antibody | 1ml x1 |
| User Instruction | 1 |
| Plate Sealer | 2 pics |
| Zipper bag | 1 pic |
Assay Principle
This elisa kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The
plate has been pre-coated with Rat TBARS antibody. TBARS present in
the sample is added and binds to antibodies coated on the wells.
And then biotinylated Rat TBARS Antibody is added and binds to
TBARS in the sample. Then Streptavidin-HRP is added and binds to
the Biotinylated TBARS 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 Rat TBARS. The reaction is terminated by addition of
acidic stop solution and absorbance is measured at 450 nm.
Precautions
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (128nmol/ml) with 120μl of
standard diluent to generate a 64nmol/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 (64nmol/ml) 1:2 with standard
diluent to produce 32nmol/ml, 16nmol/ml, 8nmol/ml and 4nmol/ml
solutions. Standard diluent serves as the zero standard(0 nmol/ml).
Any remaining solution should be frozen at -20°C and used within
one month. Dilution of standard solutions suggested are as follows:
| 64nmol/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 32nmol/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 16nmol/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 8nmol/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 4nmol/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 |
| 128nmol/ml | 64nmol/ml | 32nmol/ml | 16nmol/ml | 8nmol/ml | 4nmol/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.
