Monkey PCSK9 (Proprotein convertase subtilisin/kexin type 10) QuickTest ELISA Kit

Mfr.No.

QT-EMK0248

Sensitivity

0.375ng/ml

Detection Range

0.625-40ng/ml

Reactivity

Monkey

Assay Duration

120 minutes

Assay Type

Sandwich

Description

This kit was based on sandwich ELISA method. The experiment lasted 120 minutes. Capture antibody was conjugated to an affinity tag that was recognized by a specific antibody coated on the QuickTest plate. Add the Cap/Det Ab working solution into each well, then add the standards and pilot samples into individual wells. If the sample contains PCSK9, a capture antibody-PCSK9-biotin-detection antibody complex was formed. After incubation, unbound conjugates were removed by wash buffer. HRP-Streptavidin was added. After washing, TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. The concentration of PCSK9 in the sample was calculated by drawing a standard curve. The concentration of the target substance is proportional to the OD450 value.

Alternative Names

Proprotein convertase subtilisin/kexin type 9 ELISA Kit, Proprotein convertase 9 ELISA Kit, Subtilisin/kexin-like protease PC9 ELISA Kit, PCSK9 ELISA Kit

Detection Method

Double Antibody

Recovery

Add a certain amount of PCSK9 into the sample. Calculate the recovery by comparing the measured value with the expected amount of PCSK9 in the sample.

Sample Type	Recovery Range(%)	Average(%)
serum(n=10)	91-103	96
EDTA plasma(n=10)	87-101	96
Heparin plasma(n=10)	87-105	97

Linearity

Dilute the sample with a certain amount of PCSK9 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
serum(n=10)	90-101%	88-99%	80-96%
EDTA plasma(n=10)	86-101%	84-92%	86-94%
Heparin plasma(n=10)	94-104%	91-99%	81-97%

Specificity

Specifically binds with PCSK9 , no obvious cross reaction with other analogues.

Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.) Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(ng/ml)	OD-1	OD-2	Average
0	0.079	0.081	0.08
0.625	0.18	0.178	0.179
1.25	0.251	0.246	0.249
2.5	0.478	0.474	0.458
5	0.754	0.749	0.715
10	1.11	1.119	1.141
20	1.763	1.779	1.733
40	2.174	2.121	2.137

Assay Procedure Summary

Step 1:
Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37°C.

Washing:
Wash the plate twice without immersion.

Step 2:
Add 100ul HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37°C.

Washing:
Wash the plate five times without immersion.

Step 3:
Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)

Step 4:
Add 50ul stop solution. Read at 450nm immediately and calculate.

Precision

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/ml)	1.16	5.08	21.4	1.21	4.96	19.28
Standard deviation	0.04	0.21	0.77	0.05	0.2	0.75
CV(%)	3.61	4.08	3.61	3.84	4	3.88

Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5)	37°C for 1 month	2-8°C for 6 months
Average(%)	80	95-100

Usage Notes

Microplate reader (wavelength: 450nm)
37°C incubator (CO2 incubator for cell culture is not recommenced.)
Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
Precision single (0.5-10uL, 5-50uL, 20-200uL, 200-1000uL) and multi-channel pipette with disposable tips(Calibration is required before use.)
Sterile tubes and Eppendorf tubes with disposable tips
Absorbent paper and loading slot
Deionized or distilled water

Protocol

QT-EMK0248.pdf
MSDS

UniProt ID	A0A0D9S762

Storage	2-8°C(Sealed), Don't cryopreserve.

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SKU	Supplier No.	Size	Your price	Quantity
BHE10811345	QT-EMK0248	48T	$350		Add to Cart
BHE10811345	QT-EMK0248	96T	$500		Add to Cart

Monkey PCSK9 (Proprotein convertase subtilisin/kexin type 10) QuickTest ELISA Kit

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