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技術(shù)文章

人可溶性CD26(SCD26)ELISA試劑盒說明書

閱讀:787發(fā)布時間:2016-11-23

人可溶性CD26(SCD26)ELISA試劑盒說明書
FOR RESEARCH USE ONLY   
 
Assay range:0.5pg/ml-35pg/ml                96 determinations 
Purpose 
This kit allows for the determination of SCD26 concentrations in Human serum, cell culture supernates and 
other biological fluids 
 
Principle of the assay 
The kit assay Human SCD26 level in the sample, use 
Purified Human SCD26 antibody to coat microtiter plate wells, 
make solid-phase antibody, then add SCD26 to wells, Combined 
SCD26 antibody which With HRP labeled, become antibody - 
antigen - enzyme-antibody complex, after washing Compley, 
Add TMB substrate solution,TMB substrate becomes blue color At 
HRP enzyme-catalyzed, reaction is terminated by the addition of 
a sulphuric acid solution and the color change is measured 
spectrophotometrically at a wavelength of 450 nm. The 
concentration of Human SCD26 in the samples is then 
determined by comparing the O.D. of the samples to the standard 
curve. 

 
Materials provided with the kit 
1 wash  solution 
20ml×1bott
le 
7 Stop Solution 
6ml×1 
bottle 

HRP-Conjugate 
reagent 
6ml×1 
bottle 

Standard
(32pg/ml) 
0.5ml×1 
bottle 

Microelisa 
stripplate 
12well×8str
ips 

Standard 
diluent 
1.5ml×1bot
tle 
4 Sample diluent 
6ml×1 
bottle 
1

Instruction 1 

Chromogen 
Solution A 
6ml×1 
bottle 
1

Closure plate 
membrane 


Chromogen 
Solution B 
6ml×1 
bottle 
1

Sealed bags 1 
Specimen requirements 
1. extract as soon as possible after Specimen collection,and 
according to the relevant literature, and should be experiment 
as soon as possible after the extraction. If it can’t, specimen can 
be kept in -20 ℃ to preserve, Avoid repeated freeze-thaw 
cycles. 
2. Can’t detect the sample which contain NaN3, because NaN3 
inhibits HRP active. 
人可溶性CD26(SCD26)ELISA試劑盒說明書Assay procedure 

 
1. Dilute and add sample:Dilute Original density Standard as follow 
table: 
16pg/ml 
5 Standard 150µl Original density Standard+150µl Standard 
diluent 
8pg/ml 
4 Standard 150µl 5 Standard+150µl Standard diluent 
4pg/ml 
3 Standard 150µl 4 Standard+150µl Standard diluent 
2pg/ml 
2 Standard 150µl 3 Standard +150µl Standard diluent 
1pg/ml 
1 Standard 150µl 2 Standard +150µl Standard diluent 
2. Add sample: Set blank wells separay (blank comparison 
wells don’t add sample and HRP-Conjugate reagent, other each 
step operation is same). testing sample well. add standard 50µl 
in the Microelisa stripplate accuray,add Sample dilution 
40µl to testing sample well, then add testing sample 10µl 
(sample final dilution is 5-fold), add sample to wells , don’t 
touch the well wall as far as possible, and Gently mix. 
3. Incubate: After closing plate with Closure plate 
membrane ,incubate for 30 min at 37℃. 
4. Configurate liquid: 30-fold(or 20-fold) wash solution diluted 
30-fold (or 20-fold) with distilled water and reserve. 
5. Washing: Uncover Closure plate membrane, discard Liquid, 
dry by swing, add washing buffer to every well, still for 30s then 
drain, repeat 5 times, dry by pat. 

 
6. Add enzyme: Add HRP-Conjugate reagent 50µl to each well, 
except  blank well.  
7. Incubate: Operation with 3. 
8. Washing: Operation with 5. 
9. Color:Add Chromogen Solution A 50ul and Chromogen 
Solution B 50ul to each well, evade the light preservation for 
10 min at 37℃ 
10.Stop the reaction:Add Stop Solution50µl to each well, Stop the 
reaction(the blue color change to yellow color). 
11. Assay:take blank well as zero , Read absorbance at 450nm 
after Adding Stop Solution and within 15min. 
Steps description 
Standard, Sample 
diluent 
 
 
Add Standard, Sample diluent, incubate for 30 min at 
37℃. 
 
 
Wash 5 time,Add HRP-Conjugate reagent, incubate for 30 min at 
37℃. 

 
 
 
Wash 5 times,Add Chromogen Solution A and B, incubate for 10 min 
at 37℃. 
 
 
Add Stop Solution 
 
 
Read absorbance at 450nm within 15 min 
 
 
calculate 
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 line regression 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 factor, the result is the sample actual 

 
density. 
人可溶性CD26(SCD26)ELISA試劑盒說明書Important notes 
1. The kit takes out from the refrigeration environment should be 
balanced 15-30 minutes in the room temperature, ELISA 
plates coated if has not use up after opened, the plate should be 
stored in Sealed bag. 
2. washing buffer will Crystallization separation, it can be 
heated the water helps dissolve when dilute . Washing does not 
affect the result. 
3. 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 . 
4. if the testing material content is excessively higher (The 
sample OD is bigger than the first standard well ),please dilute 
Sample (n-fold), Please diluente and multiplied by the 
dilution factor.(×n×5). 
5. Closure plate membrane only limits the disposable use, to 
avoid cross-contamination. 
6. The substrate evade the light preservation. 
7. Please according to use instruction strictly, The test result 
determination must take the microtiter plate reader as a 

 
standard. 
8. All samples, washing buffer and each kind of reject should 
according to infective material process. 
9. Do not mix reagents with those from other lots. 
 
Storage and validity 
1.Storage:  2-8℃. 
2.validity: six months 
 


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