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Human diphtheriae antibody (C.diphtheriae-Ab) ELISA kit instruction manual

 Last Update: 2022-08-30
 Source: Internet
 Author: User

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Human Diphtheria Antibody (C.
diphtheriae-Ab) ELISA Kit Instructions

Detection principle Sample collection, processing and preservation methods Self-provided items

Microplate reader (450nm)
High-precision sampler and pipette tip: 0.
5-10uL, 2-20uL, 20-200uL, 200-1000uL
37°C incubator

Operation Precautions

The kit was stored at 2-8°C and equilibrated at room temperature for 20 minutes before use
.
There will be crystals in the concentrated washing liquid taken out from the refrigerator, which is a normal phenomenon, and the crystals are completely dissolved by heating in a water bath before use
.
The slats not used in the experiment should be immediately put back into the ziplock bag and stored in an airtight (low temperature drying)
.
The pretreated sample should be properly diluted with sample diluent according to the operation steps to achieve the best detection effect of the kit
.
Incubate in strict accordance with the time, volume and sequence indicated in the instructions
.
Shake all liquid components well before use
.

Kit composition

name	96-well configuration	48 well configuration	Remark
microtiter plate	12 holes × 8 strips	12 holes × 4 strips	none
negative control	0. 5mL	0. 5mL	none
positive control	0. 5mL	0. 5mL	none
Detection of antigen-HRP	10mL	5mL	none
20x wash buffer	25mL	15mL	Dilute according to instructions
Sample Diluent	6mL	3mL	none
Substrate A	6mL	3mL	none
Substrate B	6mL	3mL	none
Stop solution	6mL	3mL	none
Sealing film	2 sheets	2 sheets	none
manual	1 serving	1 serving	none
Ziplock bag	1	1	none

Reagent preparation and plate washing method

Manual plate washing: shake off the liquid in the wells, fill each well with washing liquid, let stand for 1 min, shake off the liquid in the wells, pat dry on absorbent paper, and wash the plate 5 times
.
Automatic plate washer: inject 350 μL of washing solution into each well, soak for 1 min, and wash the plate 5 times
.

Steps

Take out the required slats from the aluminum foil bag after equilibrating at room temperature for 20 min, and seal the remaining slats in a ziplock bag and put them back at 4°C
.
Set negative and positive control wells and sample wells, and add 50 μL of negative control and positive control to each of the negative and positive control wells;
First add 10 μL of the sample to be tested, and then add 40 μL of the sample diluent to the well of the sample to be tested;
Subsequently, 100 μL of horseradish peroxidase (HRP)-labeled detection antigen was added to each well of the negative and positive control wells and sample wells, and the reaction wells were sealed with a sealing film, and incubated at 37°C in a water bath or incubator for 60 min
.
Discard the liquid, pat dry on absorbent paper, fill each well with washing solution, let stand for 1 min, shake off the washing solution, pat dry on absorbent paper, and repeat the plate washing 5 times (you can also use a plate washer to wash the plate)
.
Add 50 μL each of substrate A and B to each well, and incubate at 37°C for 15 min in the dark
.
Add 50 μL of stop solution to each well, and measure the OD value of each well at a wavelength of 450 nm within 15 min
.

The results judge the performance of the kit

Accuracy: The average OD value of the positive control wells is ≥1.
00; the average OD value of the negative control wells is ≤0.
15, indicating that the test results are valid
.
Specificity: Does not cross-react with other soluble structural analogs
.
Repeatability: The coefficient of variation within and between plates is less than 15%
.
Storage: 2-8℃, keep away from light and moisture
.
Validity: 6 months

Disclaimer

The kit is for research use only, and shall not be used in clinical experiments or human experiments.
Otherwise, the experimenter shall bear all the consequences, and the company shall not be responsible
.

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