PCK1 Recombinant Rabbit Monoclonal Antibody [JU84-39]
Catalog# ET7106-81
PCK1 Recombinant Rabbit Monoclonal Antibody [JU84-39]
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IF-Cell
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IHC-P
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WB
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Human
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unconjugated
Overview
Product Name
PCK1 Recombinant Rabbit Monoclonal Antibody [JU84-39]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human PCK1 aa 26-75 / 622.
Species Reactivity
Human
Validated Applications
IF-Cell, IHC-P, WB
Target Molecular Weight
Predicted band size: 69 kDa
Positive Control
293T, HepG2, SH-SY5Y, human liver tissue, human kidney tissue, Hep G2 (Human hepatocellular carcinoma cell) cell lysates.
Conjugation
unconjugated
Clone Number
JU84-39
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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IF-Cell
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1:50-1:100
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IHC-P
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1:200
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WB
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1:5,000
Target
Function
Normal adjustment to changes in blood glucose levels depends on insulin signaling as well as enzymes involved in the regulation of gluconeogenesis. Pathological changes to this process are central to the type 2 diabetes phenotype. Phosphoenolpyruvate carboxykinase (PEPCK) plays an important role in this process by stimulating hepatic glucose production. PEPCK expression increases in response to glucagon and glucocorticoids, while insulin suppresses expression. Modulation of the signals governing PEPCK levels present a potential therapeutic approach to the treatment of insulin resistance and consequently obesity. The cytosolic form of PEPCK, known as PEPCK-C, and the mitochondrial form, known as PEPCK-M, are encoded by two different nuclear genes in mouse, human and chicken.
Background References
1. Adams D R et al. Three rare diseases in one sib pair: RAI1, PCK1, GRIN2B mutations associated with Smith-Magenis Syndrome, cytosolic PEPCK deficiency and NMDA receptor glutamate insensitivity. Mol Genet Metab 113:161-170 (2014).
2. Santra S et al. Cytosolic phosphoenolpyruvate carboxykinase deficiency presenting with acute liver failure following gastroenteritis. Mol Genet Metab 118:21-27 (2016).
Sequence Similarity
Belongs to the phosphoenolpyruvate carboxykinase [GTP] family.
Tissue Specificity
Major sites of expression are liver, kidney and adipocytes.
Post-translational Modification
Acetylated. Lysine acetylation by p300/EP300 is increased on high glucose conditions. Lysine acetylation promotes ubiquitination by UBR5. Acetylation is enhanced in the presence of BAG6. Deacetylated by SIRT2. Deacetylation of Lys-91 is carried out by SIRT1 and depends on PCK1 phosphorylation levels.; Phosphorylated in a GSK3B-mediated pathway; phosphorylation affects the efficiency of SIRT1-mediated deacetylation, and regulates PCK1 ubiquitination and degradation.; Ubiquitination by UBR5 leads to proteasomal degradation.
Subcellular Location
Cytoplasm, cytosol, Endoplasmic reticulum.
UNIPROT
Synonyms
cytosolic [GTP] antibody
GTP antibody
PCK1 antibody
PCKGC_HUMAN antibody
PEP carboxykinase antibody
PEPCK-C antibody
PEPCK1 antibody
PEPCKC antibody
Phosphoenolpyruvate carboxykinase 1 (soluble) antibody
Phosphoenolpyruvate carboxykinase 1 antibody
ExpandImages
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Immunocytochemistry analysis of 293T cells labeling PCK1 with Rabbit anti-PCK1 antibody (ET7106-81) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PCK1 antibody (ET7106-81) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of HepG2 cells labeling PCK1 with Rabbit anti-PCK1 antibody (ET7106-81) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PCK1 antibody (ET7106-81) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of SH-SY5Y cells labeling PCK1 with Rabbit anti-PCK1 antibody (ET7106-81) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PCK1 antibody (ET7106-81) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-PCK1 antibody (ET7106-81) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-81) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PCK1 antibody (ET7106-81) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-81) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Western blot analysis of PCK1 on Hep G2 (Human hepatocellular carcinoma cell) cell lysate with Rabbit anti-PCK1 antibody (ET7106-81) at 1/5,000 dilution.
Lysates/proteins at 20 µg/Lane.
Exposure time: 12 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ET7106-81, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 69 kDa
Observed band size: 69 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"