Glucokinase Recombinant Rabbit Monoclonal Antibody [PSH28-07]
Catalog# HA724749
Glucokinase Recombinant Rabbit Monoclonal Antibody [PSH28-07]
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WB
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IHC-P
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IF-Tissue
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IP
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Mouse
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Rat
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unconjugated
Overview
Product Name
Glucokinase Recombinant Rabbit Monoclonal Antibody [PSH28-07]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within mouse Glucokinase aa 1-465.
Species Reactivity
Mouse, Rat
Validated Applications
WB, IHC-P, IF-Tissue, IP
Target Molecular Weight
Predicted band size: 52 kDa
Positive Control
Mouse liver tissue lysate, Rat liver tissue lysate.
Conjugation
unconjugated
Clone Number
PSH28-07
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | FC(Intra) | IF-Cell | IF-Tissue | IP | |
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| human |
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| mouse |
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| rat |
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Product Features
Form
Liquid
Concentration
0.5 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*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,500
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IHC-P
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1:5,000
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IF-Tissue
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1:500
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IP
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1-2μg/sample
Target
Function
Glucokinase (EC 2.7.1.2) is an enzyme that facilitates phosphorylation of glucose to glucose-6-phosphate. Glucokinase occurs in cells in the liver and pancreas of humans and most other vertebrates. In each of these organs it plays an important role in the regulation of carbohydrate metabolism by acting as a glucose sensor, triggering shifts in metabolism or cell function in response to rising or falling levels of glucose, such as occur after a meal or when fasting. Mutations of the gene for this enzyme can cause unusual forms of diabetes or hypoglycemia. Glucokinase (GK) is a hexokinase isozyme, related homologously to at least three other hexokinases.[4] All of the hexokinases can mediate phosphorylation of glucose to glucose-6-phosphate (G6P), which is the first step of both glycogen synthesis and glycolysis. However, glucokinase is coded by a separate gene and its distinctive kinetic properties allow it to serve a different set of functions. Glucokinase has a lower affinity for glucose than the other hexokinases do, and its activity is localized to a few cell types, leaving the other three hexokinases as more important preparers of glucose for glycolysis and glycogen synthesis for most tissues and organs. Because of this reduced affinity, the activity of glucokinase, under usual physiological conditions, varies substantially according to the concentration of glucose.
Background References
1. Ashcroft FM et al. Glucokinase activity in diabetes: too much of a good thing? Trends Endocrinol Metab. 2023 Feb
2. Ren Y et al. Glucokinase as an emerging anti-diabetes target and recent progress in the development of its agonists. J Enzyme Inhib Med Chem. 2022 Dec
Subcellular Location
Cytoplasm, Nucleus, Mitochondrion.
Synonyms
ATP:D-hexose 6-phosphotransferase antibody
FGQTL3 antibody
GCK antibody
GK antibody
GLK antibody
Glucokinase antibody
Hexokinase D pancreatic isozyme antibody
Hexokinase type IV antibody
Hexokinase-4 antibody
Hexokinase-D antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of Glucokinase on different lysates with Rabbit anti-Glucokinase antibody (HA724749) at 1/2,500 dilution.
Lane 1: Mouse liver tissue lysate
Lane 2: Mouse kidney tissue lysate
Lane 3: Mouse skeletal muscle tissue lysate
Lane 4: Rat liver tissue lysate
Lane 5: Rat kidney tissue lysate
Lane 6: Rat skeletal muscle tissue lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 8 seconds; ECL: K1801
Low expression: skeletal muscle (PMID: 23085254).
Low expression of Glucokinase protein in kidney is consistent with the predicted expression pattern.
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724749, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 52 kDa
Observed band size: 52 kDa -
Western blot analysis of Glucokinase on different lysates with Rabbit anti-Glucokinase antibody (HA724749) at 1/2,500 dilution.
Lane 1: His-tagged Human Glucokinase recombinant protein
Lane 2: His-tagged Human Hexokinase II recombinant protein
Lysates/proteins at 10 ng/Lane.
Exposure time: 4 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724749, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 52 kDa
Observed band size: 52 kDa -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Liver
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Liver tissue from fasted mice. At low glucose concentrations, GCK binds to GCKR, and the resulting inactive complex is recruited into the hepatocyte nucleus (PMID: 24566088, PMID: 10713097).
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724749, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
☑ Relative expression (RE)
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Kidney
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724749, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Low expression of Glucokinase protein in kidney is consistent with the predicted expression pattern. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Liver
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Liver tissue from fasted rats. At low glucose concentrations, GCK binds to GCKR, and the resulting inactive complex is recruited into the hepatocyte nucleus (PMID: 24566088, PMID: 10713097).
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724749, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
☑ Relative expression (RE)
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Kidney
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724749, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Low expression of Glucokinase protein in kidney is consistent with the predicted expression pattern. -
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Liver
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Liver tissue from fasted mice. At low glucose concentrations, GCK binds to GCKR, and the resulting inactive complex is recruited into the hepatocyte nucleus (PMID: 24566088, PMID: 10713097).
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724749, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Application: Immunofluorescence (IF-tissue)
Species: Rat
Tissue: Liver
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Liver tissue from fasted rats. At low glucose concentrations, GCK binds to GCKR, and the resulting inactive complex is recruited into the hepatocyte nucleus (PMID: 24566088, PMID: 10713097).
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724749, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Immunoprecipitation (IP)
Glucokinase was immunoprecipitated in 0.2 mg Mouse liver tissue lysate with HA724749 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724749 at 1/2,500 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: Mouse liver tissue lysate (input)
Lane 2: HA724749 IP in mouse tissue cell lysate
Lane 3: Rabbit IgG instead of HA724749 in mouse liver tissue lysate
Exposure time: 10 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724749, 1/2,500 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 52 kDa
Observed band size: 52 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"