Hexokinase 1 Recombinant Rabbit Monoclonal Antibody [ST47-05] - BSA and Azide free
Catalog# HA750168
Hexokinase 1 Recombinant Rabbit Monoclonal Antibody [ST47-05] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Hexokinase 1 Recombinant Rabbit Monoclonal Antibody [ST47-05] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human Hexokinase 1 aa 100-140.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Target Molecular Weight
Predicted band size: 102 kDa
Positive Control
MCF7 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, Hela cell lysate, 293 cell lysate, HepG2 cell lysate, C2C12, MCF7, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Conjugation
unconjugated
Clone Number
ST47-05
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
The hexokinases utilize Mg-ATP as a phosphoryl donor to catalyze the first step of intracellular glucose metabolism, the conversion of glucose to glucose-6-phosphate. Four hexokinase isoenzymes have been identified, including hexokinase I (HXK I), hexokinase II (HXK II), hexokinase III (HXK III) and hexokinase IV (HXK IV, also designated glucokinase or GCK). Hexokinases I-III each contain an N-terminal cluster of hydrophobic amino acids. Glucokinase lacks the N-terminal hydrophobic cluster. The hydrophobic cluster is thought to be necessary for membrane binding. This is substantiated by the finding that glucokinase has lower affinity for glucose than do the other hexokinases. HXK I has been shown to be expressed in brain, kidney and heart tissues as well as in hepatoma cell lines. HXK II is involved in the uptake and utilization of glucose by adipose and skeletal tissues. Of the hexokinases, HXK III has the highest affinity for glucose. Glucokinase is expressed in pancreatic beta cells where it functions as a glucose sensor, determining the “set point” for insulin secretion.
Background References
1. Vaca Jacome A.S., et al. N-terminome analysis of the human mitochondrial proteome. Proteomics 15:2519-2524(2015).
2. Burkard T.R., et al. Initial characterization of the human central proteome. BMC Syst. Biol. 5:17-17(2011).
Sequence Similarity
Belongs to the hexokinase family.
Tissue Specificity
Isoform 2: Erythrocyte specific (Ref.6). Isoform 3: Testis-specific. Isoform 4: Testis-specific.
Subcellular Location
Mitochondrion outer membrane, Cytoplasm, Membrane, Mitochondrion.
Synonyms
BB404130 antibody
Brain form hexokinase antibody
dea antibody
DrHXK1 antibody
EC 2.7.1.1 antibody
Glycolytic enzyme antibody
HEXOKIN antibody
hexokinase I antibody
Hexokinase PI antibody
Hexokinase type I antibody
ExpandImages
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This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Western blot analysis of Hexokinase 1 on different lysates with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/1,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: C2C12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 102 kDa
Observed band size: 102 kDa
Exposure time: 60 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-28) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C2C12 cells labeling Hexokinase 1 with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of MCF7 cells labeling Hexokinase 1 with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/1,000 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 (ET1609-28) at 1/1,000 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. -
This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/1,000 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 (ET1609-28) at 1/1,000 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. -
This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Hexokinase 1 antibody (ET1609-28) at 1/1,000 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 (ET1609-28) at 1/1,000 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. -
This data was developed using ET1609-28, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of MCF7 cells labeling Hexokinase 1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1609-28, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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