Overview

  • Product nameAnti-PCSK9 antibodySee all PCSK9 primary antibodies ...
  • Description
    Rabbit polyclonal to PCSK9
  • Tested applicationsWB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Mouse PCSK9.

    (Peptide available as ab32727.)

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Liver (Mouse) Kidney (Mouse) Liver (Rat) Kidney (Rat) This antibody gave a positive signal in the following cell types: PC12. It also gave a positive signal in FFPE mouse large intestine tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab31762 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 74 kDa).Can be blocked with Mouse PCSK9 peptide (ab32727).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionMay be implicated in the differentiation of cortical neurons and may play a role in cholesterol homeostasis.
  • Tissue specificityExpressed in neuro-epithelioma, colon carcinoma, hepatic and pancreatic cell lines, and in Schwann cells.
  • Involvement in diseaseDefects in PCSK9 are the cause of familial hypercholesterolemia 3 (FH3) [MIM:603776]. FH3 inheritance is autosomal dominant.
  • Sequence similaritiesBelongs to the peptidase S8 family.
    Contains 1 peptidase S8 domain.
  • Post-translational
    modifications
    The soluble zymogen undergoes autocatalytic intramolecular processing in the endoplasmic reticulum, resulting in the cleavage of its propeptide that remains associated with the secreted enzyme.
  • Cellular localizationSecreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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
    • PC 9 antibody
    • PC9 antibody
    • PCSK 9 antibody
    • PCSK9 antibody
    • PCSK9_HUMAN antibody
    • Proprotein convertase 9 antibody
    • Proprotein convertase PC9 antibody
    • Proprotein convertase subtilisin/kexin type 9 antibody
    • PSEC0052 antibody
    • Subtilisin/kexin like protease PC9 antibody
    • Subtilisin/kexin-like protease PC9 antibody
    see all

Anti-PCSK9 antibody images

  • PCSK9 was immunoprecipitated using 0.5mg Mouse Liver tissue, 5µg of Rabbit polyclonal to PCSK9 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Liver tissue lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31762.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 62kDa; PCSK9

  • All lanes : Anti-PCSK9 antibody (ab31762) at 1 µg/ml

    Lane 1 : Liver (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate
    Lane 4 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 74 kDa
    Observed band size : 62 kDa (why is the actual band size different from the predicted?)
    ab31762 detects a band of ~62 kDa in liver and kidney lysates from mouse and rat. This corresponds to the cleaved form of PCSK9. ab31762 has not detected the 74 kDa proprotein form in the lysates tested.
  • ICC/IF image of ab31762 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab31762 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab31762 staining in mouse large intestine formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31762, 5µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-PCSK9 antibody (ab31762)

ab31762 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Liver sections)
Specification Liver sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Jun 12 2009

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