Hexokinase 2 Monoclonal antibody

• 

  WB analysis using 66974-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66974-1-Ig (Hexokinase 2 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Lamin B1 Monoclonal antibody (HRP-66095) as loading control.

• 

  IHC staining of mouse skeletal muscle using 66974-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue slide using 66974-1-Ig (Hexokinase 2 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 66974-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using Hexokinase 2 antibody (66974-1-Ig, Clone: 2A11C3 ) at dilution of 1:750 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1).

• 

  FC experiment of HepG2 using 66974-1-Ig

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug Hexokinase 2 Monoclonal antibody (66974-1-Ig, Clone:2A11C3) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1)(red), or 0.25 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Hexokinase 2 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 66974-1-Ig

Clone No.2A11C3

Publications(56)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Synonyms

HK2, HK 2, Hexokinase-B, Hexokinase-2, hexokinase2

---

Tested Applications

• Positive WB detected in: HEK-293 cells, NIH/3T3 cells, LNCaP cells, HeLa cells, HepG2 cells, Jurkat cells, K-562 cells, 4T1 cells
• Positive IHC detected in: mouse skeletal muscle tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:20000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:375-1:1500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 49 publications below
• IHC: See 12 publications below
• IF: See 3 publications below

Product Information

66974-1-Ig targets Hexokinase 2 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Hexokinase 2 fusion protein Ag16895
• Full Name: hexokinase 2
• Calculated Molecular Weight: 102 kDa
• Observed Molecular Weight: 102 kDa
• GenBank Accession Number: BC021116
• Gene Symbol: Hexokinase 2
• Gene ID (NCBI): 3099
• RRID: AB_2882294
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: P52789
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Hexokinase 2 (HK2), a rate-limiting enzyme in the first step of glycolysis pathway, expresses at high level in cancer cells compared with normal cells. HK2 provides a new target for cancer therapy due to its pivotal role in tumor tumourigenic and metastatic process. HK1 is constitutively expressed in most mammalian adult tissues. HK2, however, although is abundantly expressed in embryonic tissues, is expressed at high levels only in limited number of adult tissues such as adipose, skeletal, and cardiac muscles. (PMID: 29305912, PMID: 28427443, PMID: 23911236). The antibody also recognizes Hexokinase 1.

Publications

Species	Application	Title
mouse	WB	Cell Metab; High dietary fructose promotes hepatocellular carcinoma progression by enhancing O-GlcNAcylation via microbiota-derived acetate; Authors - Peng Zhou
mouse	WB,IF	Small; A Constant Filgotinib Delivery Adhesive Platform Based on Polyethylene Glycol (PEG) Hydrogel for Accelerating Wound Healing via Restoring Macrophage Mitochondrial Homeostasis; Authors - Jian Xie
pig	WB	Emerg Microbes Infect; The regulation of cell homeostasis and antiviral innate immunity by autophagy during classical swine fever virus infection; Authors - Xiaowen Li
human	WB	Clin Transl Med; USP7 promotes non-small-cell lung cancer cell glycolysis and survival by stabilizing and activating c-Abl; Authors - Yuanming He
mouse	WB	Br J Pharmacol; HSP90β shapes the fate of Th17 cells with the help of glycolysis-controlled methylation modification; Authors - Ling Yang
mouse	WB	Acta Pharmacol Sin; Bmal1 inhibits phenotypic transformation of hepatic stellate cells in liver fibrosis via IDH1/α-KG-mediated glycolysis.; Authors - Lei Xu