Guinea pig vascular endothelial growth factor receptor elisa kit instruction manual
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This kit is used to determine the vascular endothelial growth factor receptor (VEGFR) content in guinea pig serum, plasma and related fluid samples.
Experimental principle This kit uses a double antibody sandwich assay to determine guinea pig vascular endothelial growth factor receptor (VEGFR) levels in specimens. The microporous plate was coated with purified guinea pig vascular endothelial growth factor receptor (VEGFR) antibody to prepare a solid phase antibody, and vascular endothelial growth factor receptor (VEGFR) was sequentially added to the microcapsule of the coated monoclonal antibody, followed by HRP labeling. The vascular endothelial growth factor receptor (VEGFR) antibody binds to form an antibody-antigen-enzyme-labeled antibody complex, which is thoroughly washed and then added to the substrate TMB for color development. TMB is converted to blue under the catalysis of HRP enzyme and converted to the final yellow color by the action of an acid. The color depth is positively correlated with the vascular endothelial growth factor receptor (VEGFR) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the guinea pig vascular endothelial growth factor receptor (VEGFR) concentration in the sample was calculated from a standard curve.
Kit composition
1 20 times concentrated washing solution 20ml × 1 bottle 7 Stop solution 3ml × 1 bottle
2 enzyme standard reagent 3ml × 1 bottle 8 standard (4800 ng / L) 0.5ml × 1 bottle
3 enzyme label coating plate 12 holes × 4 strips 9 standard dilutions 1.5ml × 1 bottle
4 sample diluent 3ml × 1 bottle 10 instructions 1 copy
5 developer A liquid 3ml × 1 bottle 11 sealing film 2 sheets
6 developer B liquid 3ml × 1 bottle 12 sealed bag 1
Specimen requirements
1. The specimens should be extracted as soon as possible after collection, and the extraction should be carried out according to the relevant literature. The experiment should be carried out as soon as possible after extraction. If the test cannot be performed immediately, the specimen can be stored at -20 °C, but repeated freezing and thawing should be avoided.
2. Samples containing NaN3 could not be detected because NaN3 inhibited horseradish peroxidase (HRP) activity.
Guinea pig elisa kit operating procedure Standard dilution: This kit provides one original standard, which can be diluted in a small tube according to the following chart.
Add 2150 ng/L No. 5 standard 150 μl of the original standard to 150 μl of standard dilution
1200 ng/L No. 4 Standard 150 μl of No. 5 Standard Add 150 μl Standard Diluent
600 ng/L No. 3 Standard 150 μl of No. 4 Standard Add 150 μl Standard Diluent
300 ng/L No. 2 Standard 150 μl of No. 3 Standard Add 150 μl Standard Diluent
150 ng/L No. 1 Standard 150 μl of No. 2 Standard Add 150 μl Standard Diluent
Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes.
Solution: 20 times concentrated washing solution diluted with distilled water 20 times and then washed. Carefully remove the sealing film, discard the liquid, dry, fill each well with washing liquid, let stand for 30 seconds, then discard, repeat 5 Times, shoot dry.
Add enzyme: 50 μl of enzyme labeling reagent was added to each well, except for blank wells.
Incubation: The operation is the same as 3.
Washing: The operation is the same as 5.
Color development: Add 50 μl of color developer A to each well, then add 50 μl of color developer B, gently shake and mix, and develop color at 37 ° C for 15 minutes.
Termination: 50 μl of stop solution was added to each well to terminate the reaction (in this case, the blue color turned yellow).
Measurement: The absorbance (OD value) of each well was measured sequentially with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution. 