This gene encodes ketohexokinase that catalyzes conversion of fructose to fructose-1-phosphate. The product of this gene is the first enzyme with a specialized pathway that catabolizes dietary fructose. Alternatively spliced transcript variants encoding different isoforms have been identified.
Background References
1. Gutierrez JA et al. Pharmacologic inhibition of ketohexokinase prevents fructose-induced metabolic dysfunction. Mol Metab. 2021 Jun
2. Helsley RN et al. Ketohexokinase-C regulates global protein acetylation to decrease carnitine palmitoyltransferase 1a-mediated fatty acid oxidation. J Hepatol. 2023 Jul
Western blot analysis of ketohexokinase on different lysates with Rabbit anti-ketohexokinase antibody (HA721950) at 1/2,000 dilution.
Lane 1: HepG2 cell lysate Lane 2: Human liver tissue lysate Lane 3: Human kidney tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 33 kDa Observed band size: 33 kDa
Exposure time: 11 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 (HA721950) at 1/2,000 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.
Western blot analysis of ketohexokinase on different lysates with Rabbit anti-ketohexokinase antibody (HA721950) at 1/5,000 dilution.
Lane 1: Mouse liver tissue lysate Lane 2: Mouse kidney tissue lysate Lane 3: Rat liver tissue lysate Lane 4: Rat kidney tissue lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 33 kDa Observed band size: 33 kDa
Exposure time: 5 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 (HA721950) at 1/5,000 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.
☑ Knockdown (KD)
Western blot analysis of ketohexokinase on different lysates with Rabbit anti-ketohexokinase antibody (HA721950) at 1/2,000 dilution.
Lane 1: A549-si NT cell lysate Lane 2: A549-si ketohexokinase cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 33 kDa Observed band size: 33 kDa
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721950) at 1/2,000 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.
Immunocytochemistry analysis of HepG2 cells labeling ketohexokinase with Rabbit anti-ketohexokinase antibody (HA721950) 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-ketohexokinase antibody (HA721950) 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.
Flow cytometric analysis of HepG2 cells labeling ketohexokinase.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721950, 1μg/mL) (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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