Anti-PCSK9 antibody (ab31762)
Key features and details
- Rabbit polyclonal to PCSK9
- Suitable for: WB, IP, ICC/IF, IHC-P
- Reacts with: Mouse, Rat
- Isotype: IgG
Overview
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Product name
Anti-PCSK9 antibody
See all PCSK9 primary antibodies -
Description
Rabbit polyclonal to PCSK9 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Synthetic peptide corresponding to Mouse PCSK9 aa 650 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab32727) -
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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). 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
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Function
May be implicated in the differentiation of cortical neurons and may play a role in cholesterol homeostasis. -
Tissue specificity
Expressed in neuro-epithelioma, colon carcinoma, hepatic and pancreatic cell lines, and in Schwann cells. -
Involvement in disease
Defects in PCSK9 are the cause of familial hypercholesterolemia 3 (FH3) [MIM:603776]. FH3 inheritance is autosomal dominant. -
Sequence similarities
Belongs to the peptidase S8 family.
Contains 1 peptidase S8 domain. -
Post-translational
modificationsThe 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 localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 100102 Mouse
- Entrez Gene: 298296 Rat
- SwissProt: Q80W65 Mouse
- SwissProt: P59996 Rat
- Unigene: 133268 Mouse
- Unigene: 19195 Rat
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Alternative names
- Convertase subtilisin/kexin type 9 preproprotein antibody
- FH3 antibody
- HCHOLA3 antibody
see all
Images
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Western blot - Anti-PCSK9 antibody (ab31762)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
All lanes : 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. -
Immunocytochemistry/ Immunofluorescence - Anti-PCSK9 antibody (ab31762)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCSK9 antibody (ab31762)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. -
Immunoprecipitation - Anti-PCSK9 antibody (ab31762)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
Protocols
References (9)
ab31762 has been referenced in 9 publications.
- Carreras A et al. In vivo genome and base editing of a human PCSK9 knock-in hypercholesterolemic mouse model. BMC Biol 17:4 (2019). PubMed: 30646909
- Huang M et al. PAQR3 modulates blood cholesterol level by facilitating interaction between LDLR and PCSK9. Metabolism 94:88-95 (2019). PubMed: 30831144
- Zhang G & Li Q Inflammation Induces Lipid Deposition in Kidneys by Downregulating Renal PCSK9 in Mice with Adriamycin-Induced Nephropathy. Med Sci Monit 25:5327-5335 (2019). PubMed: 31317882
- Ding Z et al. PCSK9 expression in the ischaemic heart and its relationship to infarct size, cardiac function, and development of autophagy. Cardiovasc Res 114:1738-1751 (2018). PubMed: 29800228
- Wang L et al. Inhibition of proprotein convertase subtilisin/kexin type 9 attenuates neuronal apoptosis following focal cerebral ischemia via apolipoprotein E receptor 2 downregulation in hyperlipidemic mice. Int J Mol Med 42:2098-2106 (2018). PubMed: 30066942
- Yang SH et al. Liraglutide downregulates hepatic LDL receptor and PCSK9 expression in HepG2 cells and db/db mice through a HNF-1a dependent mechanism. Cardiovasc Diabetol 17:48 (2018). PubMed: 29618348
- Schlüter KD et al. Oxidized low-density lipoprotein (oxLDL) affects load-free cell shortening of cardiomyocytes in a proprotein convertase subtilisin/kexin 9 (PCSK9)-dependent way. Basic Res Cardiol 112:63 (2017). PubMed: 28913715
- Lai Q et al. E2F1 inhibits circulating cholesterol clearance by regulating Pcsk9 expression in the liver. JCI Insight 2:N/A (2017). PubMed: 28515357
- 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). WB . PubMed: 23862065
Images
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Western blot - Anti-PCSK9 antibody (ab31762)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
All lanes : 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. -
Immunocytochemistry/ Immunofluorescence - Anti-PCSK9 antibody (ab31762)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCSK9 antibody (ab31762)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. -
Immunoprecipitation - Anti-PCSK9 antibody (ab31762)
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
