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Instruction manual for human polyomavirus (PY) ELISA kit
Purpose of use of 48T: Experimental principle kit composition | 1 | 20x concentrated wash solution | 20ml × 1 bottle | 7 | Termination solution | 3 ml × 1 bottle |
| 2 | Microplate reagents | 3 ml × 1 bottle | 8 | Positive control | 0. 5 ml × 1 bottle |
| 3 | Microplate-coated plates | 12 holes × 4 strips | 9 | Negative control | 0. 5 ml × 1 bottle |
| 4 | Sample dilution | 3 ml × 1 bottle | 10 | Instructions | 1 serving |
| 5 | Color developer A liquid | 3 ml × 1 bottle | 11 | Sealing film | 2 sheets |
| 6 | Color developer B solution | 3ml ×1/bottle | 12 | Sealed bags | 1 pcs |
Specimens require procedures
- Numbering: The sample corresponding to the microwells are numbered in order, and each plate should be set with 2 wells of negative control, 2 wells of positive control, and 1 well of blank control (blank control wells do not add samples and microplate reagents, and the rest of the steps are the same)
- Dosing: Add 50 μl
of negative control and positive control to the negative and positive control wells, respectively.
Then add 40 μl of the sample dilution to the wells of the sample to be measured, and then add 10 μl of
the sample to be measured.
Dosing adds the sample to the bottom of the microplate plate well, trying not to touch the well wall, gently shaking and mixing well, - Incubation: Incubate at 37 °C after sealing the plate with sealing membrane for 30 min
.
- Dosing: Dilute 20x concentrated washing liquid with distilled water 20x and set aside
- Washing: Carefully remove the sealing film, discard the liquid, shake dry, fill each well with washing solution, let stand for 30 seconds and discard, repeat 5 times, pat dry
. - Add enzymes: Add 50 μl of microplate reagent per well, except for blank wells
. - Incubation: Operation same as 3
. - Washing: Operation with 5
. - Color rendering: First add color developer A50μl to each well, then add color developer B50μl, gently shake and mix well, 37 °C to avoid light for 15 minutes.
- Stop: Add 50 μl of stop solution per well to terminate the reaction (at this point the blue vertical to yellow)
is added. - Assay: Zero out the blank wells, and measure the absorbance (OD value)
of each well sequentially at a wavelength of 450 nm.
The assay should be performed
within 15 min after adding the stop solution.
Calculation and result determination: Note the storage conditions and validity period
