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ketohexokinase Recombinant Rabbit Monoclonal Antibody [PSH03-19]

Usd: 385

Specification

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Catalog# HA721950

ketohexokinase Recombinant Rabbit Monoclonal Antibody [PSH03-19]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

ketohexokinase Recombinant Rabbit Monoclonal Antibody [PSH03-19]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human ketohexokinase aa 1-298 / 298.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 33 kDa

Positive Control

HepG2 cell lysate, human liver tissue lysate, human kidney tissue lysate, mouse liver tissue lysate, mouse kidney tissue lysate, rat liver tissue lysate, rat kidney tissue lysate, HepG2.

Conjugation

unconjugated

Clone Number

PSH03-19

RRID

AB_3096619

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

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

Subcellular Location

Cytoplasm, cytosol, extracellular exosome.

UNIPROT

SwissProt: P50053 Human

SwissProt: P97328 Mouse

SwissProt: Q02974 Rat

Synonyms

EC 2.7.1.3 antibody

Fructokinase antibody

Hepatic fructokinase antibody

Ketohexokinase antibody

Ketohexokinase isoform a antibody

KHK antibody

KHK_HUMAN antibody

Images

  • Western blot analysis of ketohexokinase on different lysates with Rabbit anti-ketohexokinase antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: Human liver tissue lysate<br />Lane 3: Human kidney tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 33 kDa<br /><br />Exposure time: 11 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) 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 1/2,000 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 (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse liver tissue lysate<br />Lane 2: Mouse kidney tissue lysate<br />Lane 3: Rat liver tissue lysate<br />Lane 4: Rat kidney tissue lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 33 kDa<br /><br />Exposure time: 5 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ketohexokinase on different lysates with Rabbit anti-ketohexokinase antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si ketohexokinase cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 33 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) 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

    Exposure time: 3 minutes; ECL: K1801;
    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.

  • Immunocytochemistry analysis of HepG2 cells labeling ketohexokinase with Rabbit anti-ketohexokinase antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.

    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.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721950" style="font-weight: bold;text-decoration: underline;">HA721950</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    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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