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PCSK9 Mouse Monoclonal Antibody [2F1]

Usd: 185

Specification

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Catalog# EM1701-26

PCSK9 Mouse Monoclonal Antibody [2F1]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PCSK9 Mouse Monoclonal Antibody [2F1]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein with Human PCSK9 1-201 / 692.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 74 kDa

Positive Control

HepG2 cell lysate, K-562 cell lysate, HepG2, human liver tissue, human colon caner tissue, human kidney tissue, mouse kidney tissue.

Conjugation

unconjugated

Clone Number

2F1

RRID

AB_3068753

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG2b

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:200-1:500

  • IHC-P

  • 1:200-1:500

  • FC

  • 1:500-1:1,000

Target

Function

Proprotein convertase subtilisin/kexin type 9 (PCSK9), also known as NARC-1, is a 692 amino acid protein that belongs to the peptidase S8 family and contains one peptidase S8 domain. Important in the regulation of plasma cholesterol homeostasis, PCSK9 binds to low-density lipid receptor family members LDLR, very low-density lipid receptor (VLDLR) and apolipoprotein receptor 2 (ApoER2) and promotes their degradation in intracellular acidic compartments. PCSK9 also plays a role in neuronal differentiation and apoptosis. PCSK9 is expressed in Shwann cells, neuro-epithelioma, colon carcinoma, and hepatic and pancreatic cell lines. PCSK9 levels in the brain are highest in the cerebellum during perinatal development, with ischemia causing increased levels in the adult brain. Defects in the gene encoding this protein causes the autosomal dominant disorder familial hypercholesterolemia 3 (HCHOLA3).

Background References

1. Lai Q et al. E2F1 inhibits circulating cholesterol clearance by regulating Pcsk9 expression in the liver. JCI Insight 2:10 (2017).

2. Wen S et al. Treadmill Exercise Training Modulates Hepatic Cholesterol Metabolism and Circulating PCSK9 Concentration in High-Fat-Fed Mice. J Lipids 2013:908048 (2013).

Sequence Similarity

Belongs to the peptidase S8 family.

Tissue Specificity

Expressed in neuro-epithelioma, colon carcinoma, hepatic and pancreatic cell lines, and in Schwann cells.

Post-translational Modification

Cleavage by furin and PCSK5 generates a truncated inactive protein that is unable to induce LDLR degradation.; Undergoes autocatalytic cleavage in the endoplasmic reticulum to release the propeptide from the N-terminus and the cleavage of the propeptide is strictly required for its maturation and activation. The cleaved propeptide however remains associated with the catalytic domain through non-covalent interactions, preventing potential substrates from accessing its active site. As a result, it is secreted from cells as a propeptide-containing, enzymatically inactive protein.; Phosphorylation protects the propeptide against proteolysis.

Subcellular Location

Secreted, endoplasmic reticulum,lysosome, cytoplasm.

UNIPROT

SwissProt: Q8NBP7 Human

SwissProt: Q80W65 Mouse

Synonyms

Convertase subtilisin/kexin type 9 preproprotein antibody

FH3 antibody

HCHOLA3 antibody

Hypercholesterolemia autosomal dominant 3 antibody

LDLCQ1 antibody

NARC 1 antibody

NARC-1 antibody

NARC1 antibody

Neural apoptosis regulated convertase 1 antibody

Neural apoptosis-regulated convertase 1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of PCSK9 on different lysates with Mouse anti-PCSK9 antibody (<a href="/products/EM1701-26" style="font-weight: bold;text-decoration: underline;">EM1701-26</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: K-562 cell lysate<br />Lane 3: SH-SY5Y cell lysate (negative)<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 74 kDa<br />Observed band size: 62,78 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1701-26" style="font-weight: bold;text-decoration: underline;">EM1701-26</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of PCSK9 on different lysates with Mouse anti-PCSK9 antibody (EM1701-26) at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: SH-SY5Y cell lysate (negative)

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 74 kDa
    Observed band size: 62,78 kDa

    Exposure time: 1 minute; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-26) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling PCSK9 with Mouse anti-PCSK9 antibody (<a href="/products/EM1701-26" style="font-weight: bold;text-decoration: underline;">EM1701-26</a>) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PCSK9 antibody (<a href="/products/EM1701-26" style="font-weight: bold;text-decoration: underline;">EM1701-26</a>) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling PCSK9 with Mouse anti-PCSK9 antibody (EM1701-26) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PCSK9 antibody (EM1701-26) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human colon caner tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human colon caner tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-PCSK9 antibody. Counter stained with hematoxylin.

  • Flow cytometric analysis of HepG2 cells labeling PCSK9.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/EM1701-26" style="font-weight: bold;text-decoration: underline;">EM1701-26</a>, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling PCSK9.

    Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-26, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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