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Instructions for the use of the human soluble P-selectin sP-selectin ELISA assay

 Last Update: 2022-09-14
 Source: Internet
 Author: User

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Human (Human) soluble P-selectin (sP-selectin) ELISA test kit
Instructions for use

This kit can only be used for scientific research and shall not be used for medical diagnostic testing principles Sample collection, processing and preservation methods of self-supplied items

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

Operational considerations

Store the kit at 2-8 °C and equilibrate at room temperature for 20 min
before use.
It is normal for the concentrated washing liquid taken out of the refrigerator to crystallize, and the water bath heats the crystals to completely dissolve before use
.
The slats not used in the experiment should be immediately put back into the self-sealing bag and sealed (low temperature drying) for preservation
.
Standard S0 with a concentration of 0 can be regarded as a negative control or blank; The sample has been diluted 5 times when operating according to the instructions, and the final result is multiplied by 5 to be the actual concentration
of the sample.
The incubation operation is carried out in strict accordance with the time, amount of dosing and sequence indicated in the instructions
.
Shake well before use of all liquid components
.

Kit composition

name	96-hole configuration	48-well configuration	remark
Microplate plate	12 holes × 8	12 holes × 4 strips	not
Standards	0. 3mL*6 tubes	0. 3mL*6 tubes	not
Sample dilution	6mL	3mL	not
Detection of antibodies - HRP	10mL	5mL	not
Wash buffer for 20 ×	25mL	15mL	Dilute as directed
Substrate A	6mL	3mL	not
Substrate B	6mL	3mL	not
Termination solution	6mL	3mL	not
Sealing film	2 sheets	2 sheets	not
Instructions	1 serving	1 serving	not
Self-sealing bag	1 pcs	1 pcs	not

Preparation method of reagents

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

Procedure

Remove the desired slats from the aluminum foil bag after balancing at room temperature for 20 min, and seal the remaining slats back to 4 °C
with a self-sealing bag.
Set up standard wells and sample wells, and add 50 μL of standard products with different concentrations to each standard well;
Add 10 μL of the sample to be measured in the sample wells, and then add 40 μL of the sample dilution; Blank holes are not added
.
In addition to the blank wells, 100 μL of horseradish peroxidase (HRP)-labeled detection antibody was added to each well in the standard wells and sample wells, and the reaction wells were sealed with a sealing membrane, and incubated for 60 min
in a 37 °C water bath pot or incubator.
Discard the liquid, pat dry on the absorbent paper, fill each well with washing liquid, let stand for 1min, shake off the washing liquid, pat dry on the absorbent paper, and repeat the washing plate 5 times (you can also use the washing machine to wash the plate).
Add 50 μL of substrates A and B to each well, and incubate at 37 °C to avoid light for 15 min
.
Add 50 μL of stop solution to each well, and within 15 min, determine the OD value
of each well at a wavelength of 450 nm.

Results determine kit performance

Accuracy: The coefficient R value of the standard linear regression and the expected concentration, greater than or equal to 0.
9900
.
Sensitivity: Minimum detection concentration less than 0.
1 ng/mL
.
Specificity: does not cross-react
with other soluble structural analogues.
Repeatability: The coefficient of variation in and between plates is less than 15%.
Storage: 2-8 ° C, protected from light and moisture
.
Validity: 6 months

disclaimer

The kit is for research purposes only and shall not be used in clinical trials or human experiments, otherwise all consequences arising therefrom shall be borne by the experimenter and the company shall not be responsible.
Operate in strict accordance with the instructions, the experimenter violates the instructions, and the consequences are borne
by the experimenter.

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