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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96 Wells Mouse Anti-nuclear Antibody ELISA K High Sensitive for Research Use
Cat.No E0253Mo
Standard Curve Range: 1pg/ml - 400pg/ml
Sensitivity: 0.53pg/ml
Size: 96 wells
Assay Principle
This kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Mouse ANA antibody. ANA present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Mouse ANA Antibody is added and binds to ANA in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated ANA 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 ANA. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.
Reagent Provided
| Components | Quantity |
| Standard Solution (480pg/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 (30x) | 20ml x1 |
| Biotinylated Mouse ANA 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 (480pg/ml) with 120μl of standard diluent to generate a 240pg/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 (240pg/ml) 1:2 with standard diluent to produce 120pg/ml, 60pg/ml, 30pg/ml and 15pg/ml solutions. Standard diluent serves as the zero standard(0 pg/ml). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:
| 240pg/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 120pg/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 60pg/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 30pg/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 15pg/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 |
| 480pg/ml | 240pg/ml | 120pg/ml | 60pg/ml | 30pg/ml | 15pg/ml |
Wash Buffer Dilute 20ml of Wash Buffer Concentrate 30x 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.
