Dehydroepiandrosterone Sulfate ELISA 试剂盒

Allow the kit reagents to come to room temperature for 30 minutes.
We recommend that all standards and samples be run in duplicate to allow the end user to accurately determine DHEA-S concentrations.
Ensure that all samples have reached room temperature and have been diluted as appropriate prior to running them in the kit.
Assay Buffer Dilute Assay Buffer Concentrate 1:5 by adding one part of the concentrate to four parts of deion- ized water.
Once diluted this is stable at 4 °C for 3 months.
Wash Buffer Dilute Wash Buffer Concentrate 1:20 by adding one part of the concentrate to nineteen parts of deionized water.
Once diluted this is stable at room temperature for 3 months.
Standard Preparation Label test tubes #1 through #5.
Pipet 380 μL of Assay Buffer into tube #1 and 160 μL into tubes #2 to #5.
The Dehydroepiandrosterone sulfate (DHEA-S) stock solution contains an organic solvent.
Prerinse the pipet tip several times to ensure accurate delivery.
Carefully add 20 μL of the DHEA-S stock solution to tube #1 and vortex completely.
Take 40 μL of the DHEA-S solution in tube #1 and add it to tube #2 and vortex completely.
Repeat the serial dilutions for tubes #3 through #5.
The concentration of DHEA-S in tubes 1 through 5 will be 60,000, 12,000, 2,400, 480 and 96 pg/mL.
Use all Standards within 2 hours of preparation.

1. Use the plate layout sheet on the back page to aid in proper sample and standard identification. Determine the number of wells to be used and return unused wells to the foil pouch with desiccant. Seal the ziploc plate bag and store at 4ºC.
   2. Pipet 50 μL of samples or standards into wells in the plate.
   3. Pipet 75 μL of Assay Buffer into the non-specific binding (NSB) wells.
   4. Pipet 50 μL of Assay Buffer into wells to act as maximum binding wells (B0 or 0 pg/mL).
   5. Add 25 μL of the DetectX® Dehydroepiandrosterone sulfate Conjugate to each well using a repeater pipet.
   6. Add 25 μL of the DetectX® Dehydroepiandrosterone sulfate Antibody to each well, except the NSB wells, using a repeater pipet.
   7. Gently tap the sides of the plate to ensure adequate mixing of the reagents. Cover the plate with the plate sealer and shake at room temperature for 2 hours. If the plate is not shaken signals bound will be approximately 20 % lower.
   8. Aspirate the plate and wash each well 4 times with 300 μL wash buffer. Tap the plate dry on clean absorbent towels.
   9. Add 100 μL of the TMB Substrate to each well, using a repeater pipet.
   10. Incubate the plate at room temperature for 30 minutes without shaking.
   11. Add 50 μL of the Stop Solution to each well, using a repeater pipet.
   12. Read the optical density generated from each well in a plate reader capable of reading at 450 nm.
   13. Use the plate reader's built-in 4PLC software capabilities to calculate Dehydroepiandrosterone sulfate concentration for each sample.

Average the duplicate OD readings for each standard and sample.
Create a standard curve by reducing the data using the 4PLC fitting routine on the plate reader, after subtracting the mean OD's for the NSB.
The sample concentrations obtained, calculated from the %B/B0 curve, should be multiplied by the dilution factor to obtain neat sample values.
Or, use the MyAssays™ online tool
typical data Sample Mean OD Net OD % B/B0 Dehydroepiandrosterone sulfate (DHEA-S) Conc. (pg/mL) NSB 0.065 0 - - Standard 1 0.141 0.076 10.9 60,000 Standard 2 0.266 0.201 28.7 12,000 Standard 3 0.463 0.398 56.9 2,400 Standard 4 0.641 0.576 82.3 480 Standard 5 0.725 0.660 94.3 96 B0 0.765 0.700 100 0 Sample 1 0.308 0.243 34.7 8,208 Sample 2 0.413 0.348 49.6 3,580 Always run your own standard curve for calculation of results.
Do not use this data.
Conversion Factor: 100 pg/mL of Dehydroepiandrosterone sulfate is equivalent to 256.1 pM.