Human Proprotein Convertase 9/PCSK9 ELISA Kit - Quantikine Best Seller

Name: Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
Brand: R&D Systems
SKU: DPC900
Rating: 5 (7 reviews)

R&D Systems, part of Bio-Techne | Catalog # DPC900

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Citations (96)

Product Datasheet / COA / SDS

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Product Summary
Product Specifications
Sample Values
Precision
Recovery
Linearity
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

0.219 ng/mL

Assay Range

0.625-40 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)

View all Fibroblast Activation Protein alpha/FAP ELISA Kits »

Product Summary for Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

The Quantikine Human PCSK9 immunoassay is a 4.5 hour solid phase ELISA designed to measure PCSK9 in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant human PCSK9 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural PCSK9 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring PCSK9.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human PCSK9. This assay recognizes free and LDLR-bound PCSK9.

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human PCSK9 in this assay. No medical histories were available for the donors used in this study.

Sample Type	Mean (ng/mL)	Range (ng/mL)	Standard Deviation (ng/mL)
Serum (n=37)	313	177-460	71.5
EDTA plasma (n=37)	308	160-521	73.3
Heparin plasma (n=37)	315	159-547	81.7

Cell Culture Supernates - HepG2 human hepatocellular carcinoma cells were cultured in DMEM supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed for human PCSK9, and measured 47.2 ng/mL.

Cell Lysates - HepG2 human hepatocellular carcinoma cells were prepared as described in the Cell Lysis Procedure. An aliquot of the cell lysate was removed, assayed for human PCSK9, and measured 4.74 ng/mg of total protein.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, Cell Lysates, EDTA Plasma, Heparin Plasma, Serum

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	40	40	40
Mean (ng/mL)	4.82	14.0	27.7	4.64	14.5	27.9
Standard Deviation	0.196	0.795	1.81	0.276	0.629	1.13
CV%	4.1	5.7	6.5	6.0	4.3	4.1

Recovery for Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

The recovery of PCSK9 spiked to levels throughout the range of the assay was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Media (n=4)	107	100-111

Linearity

To assess the linearity of the assay, samples containing high concentrations of PCSK9 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Scientific Data Images for Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

Human Proprotein Convertase 9/PCSK9 ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Fibroblast Activation Protein alpha/FAP

Proprotein convertase subtilisin kexin 9 (PCSK9), also named neural apoptosis-regulated convertase 1 (NARC-1), is a member of the proteinase K subfamily of subtilisin-related serine endoproteases. The full-length protein has 692 amino acids, including a signal peptide, a pro- domain, and a catalytic domain. PCSK9 is highly expressed in the liver, intestine, and kidney. It is initially synthesized as a soluble 74 kDa precursor protein. In the endoplasmic reticulum, it undergoes autocatalytic intramolecular cleavage to generate a 14 kDa pro- domain and a 60 kDa catalytic domain. These two domains remain associated when PCSK9 is secreted outside the cells (1-3). The primary physiologic function of PCSK9 is to mediate the degradation of low density lipoprotein receptor (LDL R). Early observations indicated that gain-of-function missense mutations in the PCSK9 gene can cause an autosomal dominant form of hypercholesterolemia (4, 5). The expression of PCSK9 was observed to be up-regulated by the sterol regulatory element binding proteins (SREBPs), a family of transcription factors that are responsible for the upregulation of genes involved in cholesterol and fatty acid metabolism, such as the LDL R gene (6, 7). Further experimental evidence revealed that in mice, when the PCSK9 gene was knocked out, the number of LDL R in hepatocytes increased, whereas when PCSK9 was over-expressed, the amount of LDL R protein was reduced in the liver (8, 9). In humans, genetic analyses have shown that individuals who have nonsense or loss-of-function mutations in the PCSK9 gene have significantly lower plasma LDL cholesterol levels (10, 11). These investigations clearly indicated that PCSK9 plays a key role in reducing the hepatic LDL R levels. Recently, the underlying mechanism has been uncovered: under normal physiologic conditions, the LDL R is internalized on the cell surface and directed to the endosomes in order to be recycled back to the cell surface. PCSK9 binds to the EGF domain of the LDL R and prevents LDL R from being sorted to the endosomes. Instead, the PCSK9/LDL R complex is redistributed to the lysosomes for degradation (12-14). As such, PCSK9 regulates the amount of LDL R in the circulation and modulates cholesterol levels. Serum PCSK9 concentrations have been found to be directly associated with cholesterol levels (15, 16). Since individuals with loss-of-function PCSK9 mutations have strikingly reduced risk of coronary heart diseases, PCSK9 has become an attractive drug target in recent years (17, 18). One approach is to generate small molecules that are able to interfere with PCSK9 autoactivation and its interaction with LDL R. Other approaches aiming to reduce the amounts of PCSK9 in the circulation, such as small interfering RNAs (siRNAs), have also shown promise (19, 20).

Alternate Names

FAP, Seprase

Entrez Gene IDs

2191 (Human); 14089 (Mouse); 102134935 (Cynomolgus Monkey)

Gene Symbol

FAP

Additional Fibroblast Activation Protein alpha/FAP Products

Product Documents for Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

Product Datasheets

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COA

SDS

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Product Specific Notices for Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

For research use only

Citations
Reviews

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.