Payment & Shipping Terms:
|Discount:||Available||Lead Time:||Within 48 Hours|
|Assay Principle:||Sandwich||Sample:||Serum,plasma,urine,tissue,cell Culture Supernatant|
96 Well Plate Chicken Customized Non-ester Fatty Acid ELISA Assay Kit 2 Hours Assay Length
Standard Curve Range: 0.5nmol/ml - 200nmol/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.
This elisa test kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Chicken NEFA antibody. NEFA present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Chicken NEFA Antibody is added and binds to NEFA in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated NEFAantibody. 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 Chicken NEFA. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.
|Standard Solution (240nmol/ml)||0.5ml x1|
|Pre-coated ELISA Plate||12 * 8 well strips x1|
|Standard Diluent||3ml x1|
|Stop Solution||6ml x1|
|Substrate Solution A||6ml x1|
|Substrate Solution B||6ml x1|
|Wash Buffer Concentrate (25x)||20ml x1|
|Biotinylated Chicken NEFA Antibody||1ml x1|
|Plate Sealer||2 pics|
|Zipper bag||1 pic|
Material Required But Not Supplied
*Sample can't be diluted with this kit. Owing to the the material we use to prepare the kit, the sample matrix interference may falsely depress the specificity and accuracy of the assay.
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (240nmol/ml) with 120μl of standard diluent to generate a 120nmol/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 (120nmol/ml) 1:2 with standard diluent to produce 60nmol/ml, 30nmol/ml, 15nmol/ml and 7.5nmol/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:
|120nmol/ml||Standard No.5||120μl Original Standard + 120μl Standard Diluent|
|60nmol/ml||Standard No.4||120μl Standard No.5 + 120μl Standard Diluent|
|30nmol/ml||Standard No.3||120μl Standard No.4 + 120μl Standard Diluent|
|15nmol/ml||Standard No.2||120μl Standard No.3 + 120μl Standard Diluent|
|7.5nmol/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|
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.
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.
Contact Person: Lee