Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR15847 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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 |
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WB |
1/2500. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/1000.
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IP |
1/100.
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Notes |
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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
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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. - Information by UniProt
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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
see all -
Alternative names
- EC 2.7.1.3 antibody
- Fructokinase antibody
- Hepatic fructokinase antibody
see all
Images
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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 kDaLanes 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.
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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. -
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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
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