Hexokinase II Antibody - #DF6176

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Product: Hexokinase II Antibody
Catalog: DF6176
Description: Rabbit polyclonal antibody to Hexokinase II
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Dog, Chicken, Xenopus
Mol.Wt.: 102kDa; 102kD(Calculated).
Uniprot: P52789
RRID: AB_2838143

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MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:400
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Dog(100%), Chicken(100%), Xenopus(80%)
Clonality:
Polyclonal
Specificity:
Hexokinase II Antibody detects endogenous levels of total Hexokinase II.
RRID:
AB_2838143
Cite Format: Affinity Biosciences Cat# DF6176, RRID:AB_2838143.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

DKFZp686M1669; Hexokinase 2; Hexokinase 2 muscle; Hexokinase type II; Hexokinase-2; HK 2; HK II; HK2; HKII; HxK 2; HxK2; HXK2_HUMAN; Muscle form hexokinase;

Immunogens

Immunogen:
Uniprot:

P52789(HXK2_HUMAN) >>Visit HPA database.

Gene(ID):
HK2(3099)
Expression:
P52789 HXK2_HUMAN:

Predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle.

Description:
Hexokinase catalyzes the conversion of glucose to glucose-6-phosphate, the first step in glycolysis. Four distinct mammalian hexokinase isoforms, designated as hexokinase I, II, III, and IV (glucokinase), have been identified. Hexokinases I, II, and III are associated with the outer mitochondrial membrane and are critical for maintaining an elevated rate of aerobic glycolysis in cancer cells (Warburg Effect) (1) in order to compensate for the increased energy demands associated with rapid cell growth and proliferation (2,3).
Sequence:
MIASHLLAYFFTELNHDQVQKVDQYLYHMRLSDETLLEISKRFRKEMEKGLGATTHPTAAVKMLPTFVRSTPDGTEHGEFLALDLGGTNFRVLWVKVTDNGLQKVEMENQIYAIPEDIMRGSGTQLFDHIAECLANFMDKLQIKDKKLPLGFTFSFPCHQTKLDESFLVSWTKGFKSSGVEGRDVVALIRKAIQRRGDFDIDIVAVVNDTVGTMMTCGYDDHNCEIGLIVGTGSNACYMEEMRHIDMVEGDEGRMCINMEWGAFGDDGSLNDIRTEFDQEIDMGSLNPGKQLFEKMISGMYMGELVRLILVKMAKEELLFGGKLSPELLNTGRFETKDISDIEGEKDGIRKAREVLMRLGLDPTQEDCVATHRICQIVSTRSASLCAATLAAVLQRIKENKGEERLRSTIGVDGSVYKKHPHFAKRLHKTVRRLVPGCDVRFLRSEDGSGKGAAMVTAVAYRLADQHRARQKTLEHLQLSHDQLLEVKRRMKVEMERGLSKETHASAPVKMLPTYVCATPDGTEKGDFLALDLGGTNFRVLLVRVRNGKWGGVEMHNKIYAIPQEVMHGTGDELFDHIVQCIADFLEYMGMKGVSLPLGFTFSFPCQQNSLDESILLKWTKGFKASGCEGEDVVTLLKEAIHRREEFDLDVVAVVNDTVGTMMTCGFEDPHCEVGLIVGTGSNACYMEEMRNVELVEGEEGRMCVNMEWGAFGDNGCLDDFRTEFDVAVDELSLNPGKQRFEKMISGMYLGEIVRNILIDFTKRGLLFRGRISERLKTRGIFETKFLSQIESDCLALLQVRAILQHLGLESTCDDSIIVKEVCTVVARRAAQLCGAGMAAVVDRIRENRGLDALKVTVGVDGTLYKLHPHFAKVMHETVKDLAPKCDVSFLQSEDGSGKGAALITAVACRIREAGQR

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Chicken
100
Xenopus
80
Zebrafish
75
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P52789 As Substrate

Site PTM Type Enzyme
M1 Acetylation
Y9 Phosphorylation
K41 Ubiquitination
K49 Ubiquitination
K62 Ubiquitination
K104 Ubiquitination
Y112 Phosphorylation
K147 Ubiquitination
T153 Phosphorylation
K173 Ubiquitination
K176 Ubiquitination
Y238 Phosphorylation
K290 Ubiquitination
Y301 Phosphorylation
K315 Ubiquitination
K323 Ubiquitination
T331 Phosphorylation
K337 Ubiquitination
S340 Phosphorylation
K346 Ubiquitination
S384 Phosphorylation
K401 Ubiquitination
S415 Phosphorylation
K418 Ubiquitination
S445 Phosphorylation
S449 Phosphorylation
K451 Ubiquitination
T457 Phosphorylation
Y461 Phosphorylation
K472 Ubiquitination
T473 Phosphorylation
K488 Ubiquitination
K492 Ubiquitination
K501 Ubiquitination
S506 Phosphorylation
K510 Ubiquitination
T514 Phosphorylation
K525 Ubiquitination
K624 Ubiquitination
K638 Ubiquitination
Y686 Phosphorylation
K738 Ubiquitination
K743 Ubiquitination
S746 Phosphorylation
Y749 Phosphorylation
T762 Phosphorylation
K763 Ubiquitination
K866 Ubiquitination
K873 Ubiquitination
K885 Ubiquitination
S897 Phosphorylation
K899 Ubiquitination
T905 Phosphorylation

Research Backgrounds

Function:

Catalyzes the phosphorylation of hexose, such as D-glucose and D-fructose, to hexose 6-phosphate (D-glucose 6-phosphate and D-fructose 6-phosphate, respectively). Mediates the initial step of glycolysis by catalyzing phosphorylation of D-glucose to D-glucose 6-phosphate. Plays a key role in maintaining the integrity of the outer mitochondrial membrane by preventing the release of apoptogenic molecules from the intermembrane space and subsequent apoptosis.

Subcellular Location:

Mitochondrion outer membrane>Peripheral membrane protein. Cytoplasm>Cytosol.
Note: The mitochondrial-binding peptide (MBP) region promotes association with the mitochondrial outer membrane (PubMed:29298880). The interaction with the mitochondrial outer membrane via the mitochondrial-binding peptide (MBP) region promotes higher stability of the protein (PubMed:29298880). Release from the mitochondrial outer membrane into the cytosol induces permeability transition pore (PTP) opening and apoptosis (PubMed:18350175).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle.

Subunit Structure:

Monomer (By similarity). Interacts with TIGAR; the interaction increases hexokinase activity in a hypoxia- and HIF1A-dependent manner.

Family&Domains:

The N- and C-terminal halves of the protein contain a hexokinase domain (PubMed:29298880). In contrast to hexokinase-1 and -3 (HK1 and HK3, respectively), both hexokinase domains display catalytic activity (PubMed:29298880). The region connecting the two hexokinase domains is required for the catalytic activity of the N-terminal hexokinase domain (PubMed:29298880). The N-terminal half regulates stability of the whole enzyme (PubMed:29298880).

Belongs to the hexokinase family.

Research Fields

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Type II diabetes mellitus.

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

· Metabolism > Carbohydrate metabolism > Glycolysis / Gluconeogenesis.

· Metabolism > Carbohydrate metabolism > Fructose and mannose metabolism.

· Metabolism > Carbohydrate metabolism > Galactose metabolism.

· Metabolism > Carbohydrate metabolism > Starch and sucrose metabolism.

· Metabolism > Carbohydrate metabolism > Amino sugar and nucleotide sugar metabolism.

· Metabolism > Biosynthesis of other secondary metabolites > Neomycin, kanamycin and gentamicin biosynthesis.

· Metabolism > Global and overview maps > Metabolic pathways.

· Metabolism > Global and overview maps > Carbon metabolism.

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Digestive system > Carbohydrate digestion and absorption.

References

1). Bruceine A induces cell growth inhibition and apoptosis through PFKFB4/GSK3β signaling in pancreatic cancer. PHARMACOLOGICAL RESEARCH, 2021 (PubMed: 33992797) [IF=9.3]

Application: WB    Species: human    Sample: MIA PaCa-2 cells

Fig. 4. | Bruceine A induces cell growth inhibition and apoptosis via PFKFB4-mediated glycolysis in MIA PaCa-2 cells. (D) MIA PaCa-2 cells were treated with 25 nM bruceine A for 0, 6, 12, and 24 h. Immunoblots against GLUT1, HK2, PFKFB4,PFKM, PKM2, LDHA, and β-actin from cell lysates of MIA PaCa-2 cells were detected. β-actin was served as control. Results were expressed as means ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control cultured with 0.1% DMSO by one-way ANOVA and post hoc tests.
2). Crocetin antagonizes parthanatos in ischemic stroke via inhibiting NOX2 and preserving mitochondrial hexokinase-I. Cell death & disease, 2023 (PubMed: 36681688) [IF=9.0]

Application: WB    Species: Human    Sample: SHSY-5Y cells

Fig. 7 Crocetin reduced E3 ligase RNF146 mediated HK-I ubiquitination and proteasomal degradation. A Protein expression of HK-I in SHSY-5Y cells upon MNNG and crocetin-treatment. Cells were pretreated with concentrations of crocetin (25, 50, 100 μM) for 1 h before 4 h-MNNG (100 μM) stimulation. B Group quantification of (A) from three independent experiments. C mRNA expression of HK-I measured using qRT-PCR analysis. Cells were pretreated with concentrations of crocetin (25, 50, 100 μM) for 1 h before MNNG (100 μM) stimulation for 4 h. mRNA expression was normalized to GAPDH mRNA. D Protein expression of HK-II in SHSY-5Y cells upon MNNG and crocetin-treatment. E PAR, ubiquitination (Ub), and interaction with ubiquitin ligase RNF146 of HK-I protein in SHSY-5Y cells upon MNNG and crocetin-treatment. F–H Group quantification of Ub (F), PAR (G), and RNF146 (H) in (E). Data represent the mean ± SD from three independent experiments. I Expression of HK-I protein in SHSY-5Y cells upon RNF146-knock down (siRNA). J Group quantification of (I). Data represent the mean ± SD from three independent experiments. K Effect of RNF146 knock down (siRNA) on the PARylation and ubiquitination level of HK-I. L Docking analysis between RNF146 and PARylated HK-I. HK-I was represented in green and RNF146 shown in gray. The interaction types of residue pairs between RNF146 and HK-I were shown in detail. Significance was determined by one-way ANOVA. *p 
3). Pimozide inhibits the growth of breast cancer cells by alleviating the Warburg effect through the P53 signaling pathway. BIOMEDICINE & PHARMACOTHERAPY, 2022 (PubMed: 35658233) [IF=7.5]

Application: WB    Species: Human    Sample: MCF-7 cells

Fig. 3. Pimozide inhibits PKM2 protein and mRNA in both MCF-7 and MDA-MB-231 cells in vitro. (A-B) Cells were treated with the indicated concentrations of Pimozide for 24 h, and the protein expression of glycolytic enzymes in MCF-7(A) and MDA-MB-231(B) cells were determined by Western blot analysis (left panel). Densitometry analysis was performed to assess the glycolytic enzymes protein expression (normalized to β-actin expression), PKM2 decreased significantly compared with untreated cells (right panel). (C-D) The mRNA expression of PKM2 in MCF-7(C) and MDA-MB-231(D) cells untreated or treated with Pimozide was determined by qRT-PCR. GAPDH was used as a control. Data represent mean ± SD from three biological replicates (*p < 0.05, **p < 0.01).
4). Compound Danshen Dripping Pill inhibits doxorubicin or isoproterenol-induced cardiotoxicity. Biomedicine & Pharmacotherapy, 2021 (PubMed: 34311530) [IF=7.5]
5). Mailuo Shutong pills inhibit neuroinflammation by regulating glucose metabolism disorders to protect mice from cerebral ischemia-reperfusion injury. Journal of ethnopharmacology, 2024 (PubMed: 39053718) [IF=5.4]
6). Yes associated protein 1 promotes resistance to 5-fluorouracil in gastric cancer by regulating GLUT3-dependent glycometabolism reprogramming of tumor-associated macrophages. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2021 (PubMed: 33727040) [IF=3.9]

Application: WB    Species: Mice    Sample: GC cells

Fig. 3. IL13 secreted by YAP1-overexpressed GC cells stimulates resistance to 5-FU via inducing M2 subtype macrophage glycolysis reprogramming. A-C. RT-PCR was used to detect the mRNA expression of glycolysis enzymes, fatty acid and amino acid metabolism enzymes in THP1 after co-cultured with MKN-YAP1 or MKN45-Vetor. D. Protein level change of glycolysis enzymes in THP1 after co-cultured with MKN-YAP1, MKN45-Vetor, SGC7901-siYAP1 or SGC7901-NC. E. RT-PCR revealed the mRNA expression of glycolysis enzymes and M2 TAMs markers in THP1 after co-cultured with SGC7901-siYAP1 or SGC7901-NC. F-G. Relative lactate release from cells was determined by colorimetric analysis. Relative glucose uptake cells by Flow cytometry detection. All data presented are the mean ± SD (*p < 0.05, **p < 0.01) of triplicate determination from three independent experiments.
7). Pepsin enhances glycolysis to promote malignant transformation of vocal fold leukoplakia epithelial cells with dysplasia. European Archives of Oto-Rhino-Laryngology, 2022 (PubMed: 36380093) [IF=2.6]

Application: WB    Species: Mouse    Sample: VFL epithelial cells

Fig. 7 Efects of acidifed pepsin on Hexokinase-II (HK-II) expression and mitochondrial translocation in VFL epithelial cells. A Efects of acidifed medium (pH 3) and acidifed pepsin (pH 3, 0.1 mg/mL) on the amount of HK-II protein, as detected by Western blotting. GAPDH was served as the internal reference. On the right, strip gray values have been plotted. B Efects of acidifed medium (pH 3) and acidifed pepsin (pH 3, 0.1  mg/mL) on fuorescence expression intensity of HK-II in the mitochondria of VFL epithelial cells, as observed under a laser confocal microscope. Scale, 20 μm. One-way ANOVA was used for statistical analysis.

Application: IF/ICC    Species: Mouse    Sample: VFL epithelial cells

Fig. 7 Efects of acidifed pepsin on Hexokinase-II (HK-II) expression and mitochondrial translocation in VFL epithelial cells. A Efects of acidifed medium (pH 3) and acidifed pepsin (pH 3, 0.1 mg/mL) on the amount of HK-II protein, as detected by Western blotting. GAPDH was served as the internal reference. On the right, strip gray values have been plotted. B Efects of acidifed medium (pH 3) and acidifed pepsin (pH 3, 0.1  mg/mL) on fuorescence expression intensity of HK-II in the mitochondria of VFL epithelial cells, as observed under a laser confocal microscope. Scale, 20 μm. One-way ANOVA was used for statistical analysis.

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