Overview
- Product nameAnti-PCK2 antibodySee all PCK2 primary antibodies ...
- DescriptionRabbit polyclonal to PCK2
- Tested applicationsICC/IF, IHC-P, WB, IP more details
- Species reactivityReacts with: Rat, Human
Predicted to work with: Mouse, Pig - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human PCK2.
(Peptide available as ab87120.)
- Positive controlThis antibody gave a positive signal in HepG2 Whole Cell Lysate.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
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 | Notes |
|---|---|
| ICC/IF | ICC/IF: Use a concentration of 5 µg/ml. |
| IHC-P | IHC-P: Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| WB | WB: Use a concentration of 1 µg/ml. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa).Can be blocked with PCK2 peptide (ab87120). |
| IP | IP: Use at an assay dependent concentration. |
Target
- FunctionCatalyzes 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.
- PathwayCarbohydrate biosynthesis; gluconeogenesis.
- Involvement in diseaseDefects 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 similaritiesBelongs to the phosphoenolpyruvate carboxykinase [GTP] family.
- Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. - Cellular localizationMitochondrion.
-
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
Target information above from: UniProt accession
Q16822
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
.
-
Alternative names
- EC 4.1.1.32 antibodyGTP mitochondrial precursor antibodyHGNC:8725 antibody
- mitochondrial antibodyMitochondrial phosphoenolpyruvate carboxykinase 2 antibodyOTTHUMP00000164700 antibodyPCK2 antibodyPCKGM_HUMAN antibodyPE antibodyPEP carboxykinase antibodyPEPCK antibodyPEPCK deficiency mitochondrial antibodyPEPCK M antibodyPEPCK-M antibodyPEPCK2 antibodyPhosphoenolpyruvate carboxykinase [GTP] antibodyPhosphoenolpyruvate carboxykinase 2 (mitochondrial) antibodyPhosphoenolpyruvate carboxykinase 2 mitochondrial antibodyPhosphoenolpyruvate carboxylase antibodyPhosphopyruvate carboxylase antibody
see all
Anti-PCK2 antibody images
-
Western blot - 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. -
Immunocytochemistry/ Immunofluorescence - PCK2 antibody (ab70359)ICC/IF image of ab70359 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab70359, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa, HepG2 and Hek293 cells at 5µg/ml. -
Immunocytochemistry/ Immunofluorescence - 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.
References for Anti-PCK2 antibody (ab70359)
ab70359 has not yet been referenced specifically in any publications.
