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  1. Link

    ketohexokinase-antibody-epr15847-ab197593.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ketohexokinase antibody [EPR15847] (ab197593)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
  • Immunocytochemistry/ Immunofluorescence - Anti-ketohexokinase antibody [EPR15847] (ab197593)
  • Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
  • Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ketohexokinase antibody [EPR15847] (ab197593)
  • Immunoprecipitation - Anti-ketohexokinase antibody [EPR15847] (ab197593)
  • Anti-ketohexokinase antibody [EPR15847] (ab197593)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15847] to ketohexokinase
  • Suitable for: WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Assay
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Protein
Recombinant Human ketohexokinase protein (ab191946)

View more associated products

Overview

  • Product name

    Anti-ketohexokinase antibody [EPR15847]
    See all ketohexokinase primary antibodies
  • Description

    Rabbit monoclonal [EPR15847] to ketohexokinase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293T cell lysate; Human fetal kidney and fetal liver lysates; Mouse kidney, mouse liver, rat kidney and rat liver lysates. IHC-P: Human liver tissue. ICC/IF: HepG2 cells. IP: Human fetal kidney lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR15847
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism

Associated products

  • Alternative Versions

    • Anti-ketohexokinase antibody [EPR15847] - BSA and Azide free (ab240343)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human KHK (ketohexokinase) knockout HEK-293T cell line (ab267292)
  • KO cell lysates

    • Human KHK (ketohexokinase) knockout HEK-293T cell lysate (ab258017)
  • Recombinant Protein

    • Recombinant Human ketohexokinase protein (ab191946)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab197593 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/2500. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/100.
Notes
WB
1/2500. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/100.

Target

  • Tissue specificity

    Most abundant in liver, kidney, gut, spleen and pancreas. Low levels also found in adrenal, muscle, brain and eye.
  • Pathway

    Carbohydrate metabolism; fructose metabolism.
  • Involvement in disease

    Defects in KHK are the cause of fructosuria (FRUCT) [MIM:229800]. Fructosuria is a benign defect of intermediary metabolism.
  • Sequence similarities

    Belongs to the carbohydrate kinase pfkB family.
  • Target information above from: UniProt accession P50053 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3795 Human
    • Entrez Gene: 16548 Mouse
    • Entrez Gene: 25659 Rat
    • Omim: 614058 Human
    • SwissProt: P50053 Human
    • SwissProt: P97328 Mouse
    • SwissProt: Q02974 Rat
    • Unigene: 567297 Human
    • Unigene: 22451 Mouse
    • Unigene: 33324 Rat
    see all
  • Alternative names

    • EC 2.7.1.3 antibody
    • Fructokinase antibody
    • Hepatic fructokinase antibody
    • Ketohexokinase antibody
    • Ketohexokinase isoform a antibody
    • KHK antibody
    • KHK_HUMAN antibody
    see all

Images

  • Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    All lanes : Anti-ketohexokinase antibody [EPR15847] (ab197593) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : KHK knockout HEK293T cell lysate
    Lane 3 : Human kidney tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 33 kDa
    Observed band size: 33 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab197593 observed at 33 kDa. Red - loading control ab7291 observed at 50 kDa.

     ab197593 Anti-ketohexokinase antibody [EPR15847] was shown to specifically react with ketohexokinase in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267292 (knockout cell lysate ab258017) was used. Wild-type and ketohexokinase knockout samples were subjected to SDS-PAGE. ab197593 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Immunocytochemistry/ Immunofluorescence - Anti-ketohexokinase antibody [EPR15847] (ab197593)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling ketohexokinase with ab197593 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm and nuclear staining on HepG2 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab197593 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

  • Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    All lanes : Anti-ketohexokinase antibody [EPR15847] (ab197593) at 1/5000 dilution

    Lane 1 : Human fetal kidney lysate
    Lane 2 : Human fetal liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 33 kDa
    Observed band size: 33 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Western blot - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    All lanes : Anti-ketohexokinase antibody [EPR15847] (ab197593) at 1/2500 dilution

    Lane 1 : Mouse kidney lysate
    Lane 2 : Mouse liver lysate
    Lane 3 : Rat kidney lysate
    Lane 4 : Rat liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 33 kDa
    Observed band size: 33 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ketohexokinase antibody [EPR15847] (ab197593)

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling ketohexokinase with ab197593 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and nuclear staining on Human liver tissues is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Immunoprecipitation - Anti-ketohexokinase antibody [EPR15847] (ab197593)

    Ketohexokinase was immunoprecipitated from Human fetal kidney lysate with ab197593 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab197593 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Human fetal kidney lysate 10 µg (Input). Lane 2: ab197593 IP in Human fetal kidney lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab197593 in Human fetal kidney lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Anti-ketohexokinase antibody [EPR15847] (ab197593)
    Anti-ketohexokinase antibody [EPR15847] (ab197593)

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

Publishing research using ab197593? Please let us know so that we can cite the reference in this datasheet.

ab197593 has been referenced in 1 publication.

  • Bu P  et al. Aldolase B-Mediated Fructose Metabolism Drives Metabolic Reprogramming of Colon Cancer Liver Metastasis. Cell Metab 27:1249-1262.e4 (2018). PubMed: 29706565

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