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Human 1,25(OH)2D3 ELISA Kit
size： 96 wells
Inventory Status： In stock
This 1,25(OH)2D3 ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures. The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of 1,25(OH)2D3 in the sample, this 1,25(OH)2D3 ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus 1,25(OH)2D3 concentration. The concentration of 1,25(OH)2D3 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
|Product Name||Human 1,25(OH)2D3 ELISA Kit|
|Assay Length||2-4.5 hours|
|Assay Type||Solid Phase Sandwich ELISA|
|Validated Species||Natural and recombinant Human 1,25(OH)2D3|
|Cross Reactivity||There is no detectable cross-reactivity.|
96wells/kit, with removable strips.
|Assay Range||1.5 ng/mL – 48 ng/mL|
|Sample Type||Cell Culture Supernates, Serum，plasma,Tissue Lysates|
|Storage||Store the unopened product at 2 - 8 °C. Do not use past expiration date.|
|Sample Type||Average % Recovery||Range %|
|Cell Culture Supernates (n=10)||110||93-120|
|Tissue Lysates (n=15)||96||83-114|
|Intra-Assay Precision||Inter-Assay Precision|
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature.
The protocol for this product (see above) is intended to serve as an example only. Please refer to the Instructions For Use provided with the assay kit for precise details.
For research use only, not for use in diagnostic procedures!
1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microtiter plate.
2. Add 50μl of Standard or Sample to the appropriate wells. Blank well doesn’t add anyting.
3. Add 100μl of Enzymeconjugate to standard wells and sample wells except the blank well, cover with an adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the Microtiter Plate 4 times.
Manual Washing - Remove incubation mixture by aspirating contents of the plate into a sink or proper waste container. Using a squirt bottle, fill each well completely with Wash Solution (1X), then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure for a total of four times. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame.
Automated Washing- Aspirate all wells, then wash plates four times using Wash Buffer (1X). Always adjust your washer to aspirate as much liquid as possible and set fill volume at 350μL/well/wash. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears.
5. Add Substrate A 50μl and Substrate B 50μl to each well.Gently mix and incubate for 15 minutes at 37°C.Protect from light.
6. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.
7. Read the Optical Density (O.D.) at 450 nm using a microtiter plate readerwithin 15 minutes.
Publishing research using Human 1,25(OH)2D3 ELISA Kit? Please let us know so that we can cite the reference in this datasheet.
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