Vitamin D ELISA TEST KIT
Drug Names
Generic Name:VB12 ELISA Kit
Purpose
This kit allows for the determination of VB12 concentrations in sample.
Principle of the assay
The kit use ELISA competition to assay VD level in the sample,use Purified VD antibody
to coat microtiter plate wells, make solid-phase antibody, add VD and antibody which With HRP
labeled VD to coated microtiter wells,make them Combine competitive ,after washing
Completely, Add TMB substrate solution . the depth of color and the VD of sample were
positively correlated, measure the optical densit (OD) at 450 nm with microtiter plate reader,
calculate VD concentration by standard curve.
| Product details |
Description |
| Delivery |
Within 48 hours |
| Packaging Specifications |
8 x 12 strips, 96 wells |
| Country Of Origin |
China |
| Manufacturer |
18 months |
| Preservation method |
2℃-8℃ |
| Specimen |
Whole blood |
| Assification |
class1 |
| Type |
Elisa Test Kit |
Test principle
Materials provided with the kit
| 1 |
wash solution |
20ml×1bottle |
8
|
1 Standard(48nmol/L) |
0.5ml×1 bottle |
| 2 |
HRP-Conjugate reagent |
6ml×1 bottle |
2 Standard(24nmol/L ) |
0.5ml×1 bottle |
| 3 |
Microelisa stripplate |
12well×8strips |
3 Standard(12nmol/L) |
0.5ml×1 bottle |
| 4 |
Stop Solution |
6ml×1 bottle |
4 Standard(6nmol/L) |
0.5ml×1 bottle |
| 5 |
Chromogen Solution A |
6ml×1 bottle |
5 Standard(3nmol/L) |
0.5ml×1 bottle |
| 6 |
Chromogen Solution B |
6ml×1 bottle |
9 |
User manual |
1 |
| 7 |
Sample diluent |
6ml×1 bottle |
10 |
Closure plate membrane |
2 |
Assay procedure
1. add sample:Set blank wells separately (blank comparison wells don’t add sample and
ELISA reagent, other each step operation is same). testing sample well. add Sample
dilution 40μl to testing sample well which on ELISA plates coated, then add testing sample
10μl (sample final dilute degree is 5 times), add sample to the bottom of ELISA plates
coated well , don’t touch the well wall as far as possible, and Gently mix.
2. add enzyme:Add ELISA reagents 50μl to each well, except the blank well.
3. Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.
4. Configurate liquid: 30 times of Wash Concentrate diluted 30 times with distilled water and
reserve.
5. washing:Uncover Closure plate membrane, discard Liquid, dry by swing, add washing
buffer to every well, still for 30 second then remove, repeat 5 times, dry by pat.
6. color:add color reagent A 50μl and color reagent B 50μl to each well. Gently mix, incubate
for 10 min at 37℃.
7. Stop the reaction:Add Stop Solution50μl to each well, Stop the reaction(the blue color
change to yellow color Immediately).
8. assay:take blank well as zero , measure the optical densit (OD) at 450 nm after Adding
Stop Solution and within 15min
Calculate
Take the standard density as the horizontal, the OD value for the vertical,draw the
standard curve on graph paper, Find out the corresponding density according to the sample
OD value by the Sample curve, multiplied by the dilution multiple, or calculate the straight lineregression equation of the standard curve with the standard density and the OD value ,with
the sample OD value in the equation, calculate the sample density, multiplied by the dilution
multiple, the result is the sample actual density.
Important notes
- The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature then use, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.
- washing buffer will Crystallization separation, it can be heated the water helps dissolve when
dilute . Washing does not affect the result.
- add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 min, if the number of sample is much , recommend to use Volley .
- Please make specification curve when you assay, had better make duplicate well, if in the sample the testing material content is excessively high (The sample OD is bigger than the first standard well OD ),please use Sample dilution to dilute certain multiple (n times),then assay. Please multiply total Dilution Times when calculate(×n×5).
- Closure plate membrane only limits the disposable use, in order to avoid the overlapping pollution.
- The substrate please evade the light preservation.
- Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard .
- All samples, washing buffer and each kind of reject should according to infective material process.
- This reagent which different batch number component do not mix.
- If it’s different form English instruction, take English instruction as the standard.
Storage and stability
• Store at 2-8℃.
• Seal and return unused reagents to 2-8℃, under which conditions the stability will be retained for 2 months, or until the labeled expiry date, whichever is earlier.
