Mouse Proprotein Convertase 9 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY3985
Ancillary Products Available
Mouse Proprotein Convertase 9 / PCSK9 ELISA Standard Curve
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Product Details
Citations (2)
FAQs
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Mouse Proprotein Convertase 9 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse Proprotein Convertase 9 / PCSK9. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet ELISA.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Reagent Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product .

Data Example

Mouse Proprotein Convertase 9 / PCSK9 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. 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: Proprotein Convertase 9/PCSK9

PCSK9 (proprotein convertase subtilisin kexin 9), also called proprotein convertase 9 or NARC-1 (neural apoptosis-regulated convertase 1), is a member of the proteinase K subfamily of subtilisinrelated serine endoproteases. Mouse PCSK9 cDNA encodes 694 amino acids, including a signal peptide, a prodomain, 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 prodomain and a 60 kDa catalytic domain. While within the secretion pathway, the prodomain remains associated and functions as a chaperone for the catalytic domain (1-4). During secretion, a portion of active PCSK9 may undergo additional N-terminal proteolysis by furin or proprotein convertase 5/6A, creating an inactive 53 kDa form (5). This cleavage site is conserved between mouse and human or rat PCSK9, which share 78% or 93% amino acid sequence identity, respectively. While the 60 kDa protein is the major form, its ratio with the 53 kDa forms is variable in humans (5, 6). 

The primary physiologic function of PCSK9 is to mediate the degradation of low density lipoprotein receptor (LDLR). Early observations indicated that gain-of-function missense mutations in the human PCSK9 gene can cause an autosomal dominant form of hypercholesterolemia (7, 8). The expression of PCSK9 is also upregulated 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 LDLR gene (9, 10). Further experimental evidence revealed that when the mouse PCSK9 gene is deleted, LDLR expression in hepatocytes is increased. Conversely, PCSK9 over-expression decreases liver LDLR protein expression (11, 12). 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, while in mouse, administration of a PCSK9 neutralizing antibody or antisense oligonucleotides lowers serum cholesterol (1, 13-15). These investigations clearly indicate that PCSK9 plays a key role in reducing the hepatic LDLR levels. Paradoxically, administration of cholesterol-lowering drugs such as statins appear to enhance production of PCSK9 (6). 
The underlying mechanism of cholesterol regulation by PCSK9 is as follows: under normal physiologic conditions, the LDLR is internalized at 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 LDLR and prevents LDLR from being sorted to the endosomes. Instead, the PCSK9/LDLR complex is redistributed to the lysosomes for degradation (16-18). As such, PCSK9 regulates the amount of LDLR in the circulation and hence, modulates cholesterol levels. Serum PCSK9 concentrations have been found to be directly associated with cholesterol levels (19, 20). Since PCSK9 loss-of-function mutations strikingly reduce risk of coronary heart diseases, PCSK9 has become an attractive drug target (1, 21, 22). One approach is to generate small molecules that are able to interfere with PCSK9 autoactivation and its interaction with LDLR. Other approaches aiming to reduce the amount of PCSK9 in the circulation, such as small interfering RNAs (siRNAs), have also shown promise (23, 24).

Entrez Gene IDs:
255738 (Human); 100102 (Mouse); 298296 (Rat); 102142788 (Cynomolgus Monkey)
Alternate Names:
EC 3.4.21; EC 3.4.21.111; FH3; FH3neural apoptosis regulated convertase 1; FHCL3; HCHOLA3; hypercholesterolemia, autosomal dominant 3; LDLCQ1; NARC1; NARC-1; NARC-1convertase subtilisin/kexin type 9 preproprotein; NARC1EC 3.4.21.-; Neural apoptosis-regulated convertase 1; PC9; PCSK9; Proprotein Convertase 9; proprotein convertase subtilisin/kexin type 9; Subtilisin/kexin-like protease PC9

Citations for Mouse Proprotein Convertase 9 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Deletion of NoxO1 limits atherosclerosis development in female mice
    Authors: GK Buchmann, C Schürmann, T Warwick, MH Schulz, M Spaeth, OJ Müller, K Schröder, H Jo, N Weissmann, RP Brandes
    Redox Biol, 2020;37(0):101713.
    Species: Mouse
    Sample Types: Serum
  2. Effect of Porphyromonas gingivalis infection on post-transcriptional regulation of the low-density lipoprotein receptor in mice.
    Authors: Miyazawa, Haruna, Tabeta, Koichi, Miyauchi, Sayuri, Aoki-Nonaka, Yukari, Domon, Hisanori, Honda, Tomoyuki, Nakajima, Takako, Yamazaki, Kazuhisa
    Lipids Health Dis, 2012;11(0):121.
    Species: Mouse
    Sample Types: Serum

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