Product Name
Insect Phosphofructokinase(PFK) ELISA Kit
Product Type
Insect ELISA Kit
Description
This kit adopts double antibody sandwich ELISA method.
Biotinylated antibodies and horseradish peroxidase-labeled affinities are added sequentially.
The TMB appears blue and turns yellow after the addition of termination solution.
The OD value was measured at 450 nm, and the concentration of antigen was proportional to the OD450 value.
Use
For scientific research experiments.
Application
This kit is used for in vitro quantitative detection of concentrations in serum, plasma, or other relevant biological fluids.
Composition
Instructions;
Sealing film;
Sealed bag;
Enzyme-coated plates;
Standards;
Microplate labeling reagents;
Sample diluent;
Color developer A solution;
Color developer B solution;
Terminating solution;
20 times-Concentrate the washing liquid
Note
All reagent bottles must be tightly capped to prevent evaporation and microbial contamination.
Please measure rather than pour directly when using.
Sample Handling and Requirements
1. Serum: Blood was naturally clotted at RM for 15 minutes and centrifuged for about 20 minutes (2,000-3,000 rpm).
2. Plasma: EDTA or sodium citrate should be selected as anticoagulant, mixed for 15 minutes, the time and speed of centrifugationare the same as above.
3. Urine: Collect in sterile tubes, the time and speed of centrifugationare the same as above.
4. Cell culture supernatant: When detecting secretory components, collect in sterile tubes. The time and speed of centrifugationare the same as above.
5. Samples containing NaN3 cannot be tested because NaN3 inhibits horseradish peroxidase (HRP) activity.
Operation Step
1. Standard spiking: Add 50 μL standards to well.
2. Sample addition: Add 40 μL of sample dilution to the sample wells on the enzyme plate, and then add 10 μL of the sample to be tested (the final dilution of the sample is 5 times), and mix well.
3. Add enzyme and warming: Add 100 μL of enzyme reagent to each well, except for blank wells, 37℃ for 60 minutes.
4. Washing: Discard the liquid, add washing solution (diluted 20 times), 30 seconds and discard, repeat 5 times.
5. Color development: Add 50 μL CA and then 50 μL CB and mix, 37℃ for 15 minutes at avoiding light.
6. Determination: Measure the OD after the addition of the termination solution.
Kit Performance
1. The R value of the correlation coefficient between the sample linear regression and the expected concentration is above 0.95.
2. The intra-batch coefficient and inter-batch coefficient of variation should be less than 10% and 15%, respectively.
