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ELISA Kit for Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9)

FH3; HCHOLA3; NARC1; Hypercholesterolemia,Autosomal Dominant 3; Neural apoptosis-regulated convertase 1; Proprotein convertase 9; Subtilisin/kexin-like protease PC9

Specificity

This assay has high sensitivity and excellent specificity for detection of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9).
No significant cross-reactivity or interference between Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and the recovery rates were calculated by comparing the measured value to the expected amount of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-97 92
EDTA plasma(n=5) 91-101 95
heparin plasma(n=5) 94-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 78-103% 94-103% 90-104% 91-105%
EDTA plasma(n=5) 93-101% 87-102% 96-104% 97-105%
heparin plasma(n=5) 95-105% 92-102% 98-105% 88-95%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Biological Trace Element Research Synergistic Cardioprotective Effects of Combined Chromium Picolinate and Atorvastatin Treatment in Triton X-100-Induced Hyperlipidemia in Rats: Impact on Some Biochemical Markers pubmed:28409410
BASIC RESEARCH IN CARDIOLOGY Autocrine effects of PCSK9 on cardiomyocytes Pubmed: 33169229
J Transl Med Multi-omic signatures of atherogenic dyslipidaemia: pre-clinical target identification and validation in humans 33407555
Metabolism MiR-337-3p lowers serum LDL-C level through targeting PCSK9 in hyperlipidemic mice 33775647
Eur Rev Med Pharmacol Sci Association of serum proprotein convertase Subtilisin/Kexin Type 9 (PCSK9) level with thyroid function disorders 34533801
Clin Exp Hypertens Proprotein convertase subtilisin/kexin type 9 is associated with atherosclerosis in patients with Behcet's disease Pubmed:35502687
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