Anti-PCK2 antibody (ab70359)
Key features and details
- Rabbit polyclonal to PCK2
- Suitable for: IHC-P, WB, ICC, IP
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-PCK2 antibody
See all PCK2 primary antibodies -
Description
Rabbit polyclonal to PCK2 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC, IPmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Pig
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Immunogen
Synthetic peptide corresponding to Human PCK2 aa 400-500 conjugated to keyhole limpet haemocyanin.
(Peptide available asab87120) -
Positive control
- This antibody gave a positive signal in HepG2 Whole Cell Lysate.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
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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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab70359 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 IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. WB Use a concentration of 1 µg/ml. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa). ICC Use a concentration of 1 µg/ml. IP Use at an assay dependent concentration. Target
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Function
Catalyzes the conversion of oxaloacetate (OAA) to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle. -
Pathway
Carbohydrate biosynthesis; gluconeogenesis. -
Involvement in disease
Defects in PCK2 are the cause of mitochondrial phosphoenolpyruvate carboxykinase deficiency (M-PEPCKD) [MIM:261650]. A metabolic disorder resulting from impaired gluconeogenesis. It is a rare disease with less than 10 cases reported in the literature. Clinical characteristics include hypotonia, hepatomegaly, failure to thrive, lactic acidosis and hypoglycemia. Autoposy reveals fatty infiltration of both the liver and kidneys. The disorder is transmitted as an autosomal recessive trait. -
Sequence similarities
Belongs to the phosphoenolpyruvate carboxykinase [GTP] family. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Mitochondrion. - Information by UniProt
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Database links
- Entrez Gene: 5106 Human
- Entrez Gene: 74551 Mouse
- Entrez Gene: 361042 Rat
- Omim: 261650 Human
- SwissProt: Q16822 Human
- SwissProt: Q8BH04 Mouse
- Unigene: 75812 Human
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Alternative names
- EC 4.1.1.32 antibody
- GTP mitochondrial precursor antibody
- HGNC:8725 antibody
see all
Images
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Immunocytochemistry - Anti-PCK2 antibody (ab70359)
ab70359 staining Phosphoenolpyruvate carboxykinase [GTP], mitochondrial precursor in MCF7 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab70359 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Western blot - Anti-PCK2 antibody (ab70359)Anti-PCK2 antibody (ab70359) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 71 kDa
Additional bands at: 40 kDa, 50 kDa. We are unsure as to the identity of these extra bands. -
Immunoprecipitation - Anti-PCK2 antibody (ab70359)PCK2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to PCK2 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, HepG2 whole cell extract 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab70359.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 71kDa: PCK2. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCK2 antibody (ab70359)IHC image of PCK2 staining in normal human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab70659, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Protocols
Datasheets and documents
References (8)
ab70359 has been referenced in 8 publications.
- Zhu X et al. CREB-upregulated lncRNA MEG3 promotes hepatic gluconeogenesis by regulating miR-302a-3p-CRTC2 axis. J Cell Biochem 120:4192-4202 (2019). PubMed: 30260029
- Xiang J et al. Dietary Supplementation of Vine Tea Ameliorates Glucose and Lipid Metabolic Disorder via Akt Signaling Pathway in Diabetic Rats. Molecules 24:N/A (2019). PubMed: 31096578
- Niu L et al. Effects of chronic dexamethasone administration on hyperglycemia and insulin release in goats. J Anim Sci Biotechnol 9:26 (2018). PubMed: 29568520
- Liu MX et al. Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma. Oncogene 37:1637-1653 (2018). PubMed: 29335519
- Ren Z et al. Enhancement of porcine intramuscular fat content by overexpression of the cytosolic form of phosphoenolpyruvate carboxykinase in skeletal muscle. Sci Rep 7:43746 (2017). WB . PubMed: 28252054
- Méndez-Lucas A et al. Mitochondrial PEPCK is a Pro-Survival, ER-Stress Response Gene Involved in Tumor Cell Adaptation to Nutrient Availability. J Biol Chem N/A:N/A (2014). WB, IHC-P ; Human . PubMed: 24973213
- Méndez-Lucas A et al. PEPCK-M expression in mouse liver potentiates, not replaces, PEPCK-C mediated gluconeogenesis. J Hepatol 59:105-13 (2013). WB, ICC/IF ; Mouse, Rat, Rabbit, Human . PubMed: 23466304
- Yang Y et al. A novel fumarate hydratase-deficient HLRCC kidney cancer cell line, UOK268: a model of the Warburg effect in cancer. Cancer Genet 205:377-90 (2012). PubMed: 22867999
Images
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Immunocytochemistry - Anti-PCK2 antibody (ab70359)
ab70359 staining Phosphoenolpyruvate carboxykinase [GTP], mitochondrial precursor in MCF7 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab70359 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Western blot - Anti-PCK2 antibody (ab70359)Anti-PCK2 antibody (ab70359) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 71 kDa
Additional bands at: 40 kDa, 50 kDa. We are unsure as to the identity of these extra bands.
-
Immunoprecipitation - Anti-PCK2 antibody (ab70359)PCK2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to PCK2 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, HepG2 whole cell extract 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab70359.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 71kDa: PCK2. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCK2 antibody (ab70359)IHC image of PCK2 staining in normal human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab70659, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
