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Feline Astrovirus (Astrovirus) ELISA Kit Instructions
96T purpose of use: experimental principle kit composition| 1 | 30x concentrated washing solution | 20ml×1 bottle | 7 | Stop solution | 6ml×1 bottle |
| 2 | enzyme labeling reagent | 6ml×1 bottle | 8 | positive control | 0. 5ml×1 bottle |
| 3 | ELISA coated plate | 12 holes × 8 strips | 9 | negative control | 0. 5ml×1 bottle |
| 4 | Sample Diluent | 6ml×1 bottle | 10 | manual | 1 serving |
| 5 | Color developer liquid A | 6ml×1 bottle | 11 | Sealing film | 2 sheets |
| 6 | Color developer liquid B | 6ml×1/bottle | 12 | sealed bag | 1 |
Specimen Requirements Operation Steps
- Numbering: Number the microwells corresponding to the samples in sequence.
Each plate should have 2 wells for negative control, 2 wells for positive control, and 1 well for blank control (the blank control well does not add samples and enzyme labeling reagents, and the other steps are the same) - Add sample: Add 50μl of negative control and positive control to the negative and positive control wells respectively
.
Then add 40 μl of sample diluent to the well of the sample to be tested, and then add 10 μl of the sample to be tested
.
Add the sample and add the sample to the bottom of the well of the ELISA plate, try not to touch the wall of the well, and shake gently to mix. - Incubation: Seal the plate with a sealing film and incubate at 37°C for 30 minutes
. - Dosing: Dilute the 30-fold concentrated washing solution with distilled water 30-fold for later use
- Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with washing solution, let stand for 30 seconds, and then discard, repeat this process 5 times, and pat dry
. - Add enzyme: Add 50μl of enzyme labeling reagent to each well, except for blank wells
. - Incubation: operation is the same as 3
. - Washing: the operation is the same as 5
. - Color development: first 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 in the dark.
- Termination: Add 50 μl of stop solution to each well to stop the reaction (the blue turns to yellow at this time)
. - Determination: Zero the blank well, and measure the absorbance (OD value) of each well in sequence at a wavelength of 450 nm
.
The measurement should be carried out within 15 minutes after adding the stop solution
.
Calculations and Results Determination: Considerations
- The sealing film is for single use only to avoid cross-contamination
.
