Shanghai Korain Biotech Co., Ltd
Shanghai Korain Biotech Co Ltd
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Specificity and sensitivity Rat Ghrelin ELISA Kit 96 wells
Cat.No E0896Ra
Standard Curve Range: 40ng/L - 12000ng/L
Sensitivity: 20.56ng/L
Size: 96 wells
Storage: Store the reagents at 2-8°C. For over 6-month storage refer to the expiration date keep it at -20°C. Avoid repeated thaw cycles. If individual reagents are opened it is recommended that the kit be used within 1 month.
* This product is for research use only, not for use in diagnosis procedures. It’s highly recommend to read this instruction entirely before use.
Intended Use
This sandwich kit is for the accurate quantitative detection of Rat Ghrelin (also known as GHRL) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.
Assay Principle
This kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Rat GHRL antibody. GHRL present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Rat GHRL Antibody is added and binds to GHRL in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated GHRL 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 GHRL. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.
Reagent Provided
| Components | Quantity |
| Standard Solution (12800ng/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 Rat GHRL Antibody | 1ml x1 |
| User Instruction | 1 |
| Plate Sealer | 2 pics |
| Zipper bag | 1 pic |
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (12800ng/L) with 120μl of standard diluent to generate a 6400ng/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 (6400ng/L) 1:2 with standard diluent to produce 3200ng/L, 1600ng/L, 800ng/L and 400ng/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:
| 6400ng/L | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 3200ng/L | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 1600ng/L | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 800ng/L | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 400ng/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 |
| 12800ng/L | 6400ng/L | 3200ng/L | 1600ng/L | 800ng/L | 400ng/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.
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.
References
Ariyasu H., Takaya K., Hosoda H., Iwakura H., Ebihara K., Mori K., Ogawa Y., Hosoda K., Akamizu T., Kojima M., Kangawa K., Nakao K.
Endocrinology 143:3341-3350(2002)