Human Hexokinase 1/2 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB8179

R&D Systems, part of Bio-Techne
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MAB8179
MAB8179-SP
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Simple Western, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 927312

View all Hexokinase 1/2 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Hexokinase 2.
Phe11-Arg917
Accession # P52789

Specificity

Detects human Hexokinase 2 in direct ELISAs. Detects human Hexokinase 1 and human Hexokinase 2 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Hexokinase 1/2 Antibody

Detection of Human Hexokinase 1/2 antibody by Western Blot.

Detection of Human Hexokinase 1/2 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Hexokinase 1/2 Monoclonal Antibody (Catalog # MAB8179) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Hexokinase 1/2 at approximately 105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Hexokinase 1/2 antibody in Human Cervical Cancer Tissue by Immunohistochemistry (IHC-P).

Hexokinase 1/2 in Human Cervical Cancer Tissue.

Hexokinase 1/2 was detected in immersion fixed paraffin-embedded sections of human cervical cancer tissue using Mouse Anti-Human Hexokinase 1/2 Monoclonal Antibody (Catalog # MAB8179) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Hexokinase 1/2 antibody by Simple WesternTM.

Detection of Human Hexokinase 1/2 by Simple WesternTM.

Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Hexokinase 1/2 at approximately 96 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Hexokinase 1/2 Monoclonal Antibody (Catalog # MAB8179) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human Hexokinase 1/2 Antibody

Application
Recommended Usage

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human cervical cancer tissue

Simple Western

5 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line

Western Blot

0.2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line

Reviewed Applications

Read 1 review rated 5 using MAB8179 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Hexokinase 1/2

Hexokinases phosphorylate hexose to form hexose 6-phosphate, the first step in hexose metabolism (1). Phosphorylation of a hexose adds charge to molecule thereby making it difficult to transport out of a cell. The hexose is therefore retained for intracellular metabolic processes, such as glycolysis or glycogen synthesis. In most organisms, glucose is the most important substrate of hexokinases and glucose-6-phosphate is the most important product. There are four mammalian hexokinases (2). Hexokinase 1, 2 and 3 are referred to as high-affinity hexokinases because their Km for glucose is below 1 mM. Hexokinase 4 is specific for glucose and is also referred to as glucokinase (3). Hexokinase 2 (HK2), also known as muscle form hexokinase, localizes to the outer membrane of mitochondria and is present in adipose tissue, skeletal muscle, and heart (4). The amino acids corresponding to the mitochondrial binding domain (5) have been removed in the recombinant enzyme. Like Hexokinase 1 (HK1), HK2 contains two homologous halves that may have evolved from an ancestral hexokinase through gene duplication and tandem ligation (6). Unlike HK1, in HK2 both the C-terminal and N-terminal portions are catalytically active with the N-terminal half having higher activity than the C-terminal half (7). In HK2 both the N-terminal and C-terminal halves exhibit product inhibition. HK2 overexpression is required for tumor growth making HK2 an attractive oncotarget (4). The enzymatic activity of recombinant human HK2 is measured using a phosphatase-coupled method (8).

References

  1. Aleshin, A.E. et al. (1998) Structure 6:39-50
  2. Takeda, J. et al. (1993) J. Biol. Chem. 268:15200.
  3. Lange, A.J. et al. (1991) Biochem. J. 277: 159.
  4. Patra, K.C. et al. (2013) Cancer Cell. 24:213.
  5. Bianchi M et al. (1998) Mol Cell Biochem 189:185.
  6. Ureta, T. (1982) Comp Biochem Physiol. B. 71B:549.
  7. Ahn, K.J. et al. (2009) BMB Rep. 42:350.
  8. Wu, Z.L. (2011) PLoS One 6:e23172.

UniProt

P52789

Additional Hexokinase 1/2 Products

Product Documents for Human Hexokinase 1/2 Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Human Hexokinase 1/2 Antibody

For research use only

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