Sandwich Type Mouse Leptin Receptor ELISA Test Kit Highly Sensitive With Oem Service

Cat.No E0650Mo

Reagent Provided

Material Required But Not Supplied

• 37°C±0.5°C incubator
• Absorbent paper
• Precision pipettes and disposable pipette tips
• Clean tubes
• Deionized or distilled water
• Microplate reader with 450 ± 10nm wavelength filter

Intended Use
This sandwich kit is for the accurate quantitative detection of Mouse Leptin Receptor (also known as LR/Ob-R) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
CV(%) = SD/mean x 100
Intra-Assay: CV<8%
Inter-Assay: CV<10%

Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (32ng/ml) with 120μl of standard diluent to generate a 16ng/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 (16ng/ml) 1:2 with standard diluent to produce 8ng/ml, 4ng/ml, 2ng/ml and 1ng/ml solutions. Standard diluent serves as the zero standard(0 ng/ml). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:

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.

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.

Referances
"Characterizaton of short isoforms of the leptin receptor in rat cerebral microvessels and of brain uptake of leptin in mouse models of obesity."
Hileman S.M., Pierroz D.D., Masuzaki H., Bjoerbaek C., El-Haschimi K., Banks W.A., Flier J.S.
Endocrinology 143:775-783(2002)