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 Activin A ACV-A ELISA Kit Sensitivity and
Specificity
Cat.No E0792Mo
Standard Curve Range: 10ng/L - 3000ng/L
Sensitivity: 5.43ng/L
Size: 96 wells
Assay Principle
This kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate
has been pre-coated with Mouse ACV-A antibody. ACV-A present in the
sample is added and binds to antibodies coated on the wells. And
then biotinylated Mouse ACV-A Antibody is added and binds to ACV-A
in the sample. Then Streptavidin-HRP is added and binds to the
Biotinylated ACV-A 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 ACV-A. The reaction is terminated by addition of
acidic stop solution and absorbance is measured at 450 nm.
Reagent Provided
| Components | Quantity |
| Standard Solution (3200ng/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 (30x) | 20ml x1 |
| Biotinylated Mouse ACV-A 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 (3200ng/L) with 120μl of
standard diluent to generate a 1600ng/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 (1600ng/L) 1:2 with standard
diluent to produce 800ng/L, 400ng/L, 200ng/L and 100ng/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:
| 1600ng/L | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 800ng/L | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 400ng/L | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 200ng/L | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 100ng/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 |
| 3200ng/L | 1600ng/L | 800ng/L | 400ng/L | 200ng/L | 100ng/L |
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.
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.
Calculation of Result
Construct a standard curve by plotting the average OD for each
standard on the vertical (Y) axis against the concentration on the
horizontal (X) axis and draw a best fit curve through the points on
the graph. These calculations can be best performed with
computer-based curve-fitting software and the best fit line can be
determined by regression analysis.
