Overview

  • Product name

    Anti-Hexokinase II antibody [EPR20839]
    See all Hexokinase II primary antibodies
  • Description

    Rabbit monoclonal [EPR20839] to Hexokinase II
  • Host species

    Rabbit
  • Specificity

    Not recommended for rat and mouse tissues in IHC.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Hexokinase II aa 1-500. The exact sequence is proprietary.
    Database link: P52789

  • Positive control

    • WB: Hexokinase II recombinant protein (aa16-458); HeLa, SH-SY5Y and NIH/3T3 whole cell lysates; HKDC1 and Hexokinase I-knockdown HeLa cell lysates; Rat testis and muscle lysates; Human testis, colon and fetal heart lysates; Mouse muscle lysate. IHC-P: Human heart and pancreatic adenocarcinoma tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209847 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 1/1000. Detects a band of approximately 102 kDa (predicted molecular weight: 102 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended for rat and mouse tissues.

ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/60.

Target

  • Tissue specificity

    Predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle.
  • Pathway

    Carbohydrate metabolism; hexose metabolism.
  • Sequence similarities

    Belongs to the hexokinase family.
    Contains 2 hexokinase domains.
  • Domain

    The N- and C-terminal halves of this hexokinase show extensive sequence similarity to each other. The catalytic activity is associated with the C-terminus while regulatory function is associated with the N-terminus. Each domain can bind a single glucose and Gluc-6-P molecule.
  • Cellular localization

    Mitochondrion outer membrane. Its hydrophobic N-terminal sequence may be involved in membrane binding.
  • Information by UniProt
  • Database links

  • Alternative names

    • DKFZp686M1669 antibody
    • Hexokinase 2 antibody
    • Hexokinase 2 muscle antibody
    • Hexokinase type II antibody
    • Hexokinase-2 antibody
    • HK 2 antibody
    • HK II antibody
    • HK2 antibody
    • HKII antibody
    • HxK 2 antibody
    • HxK2 antibody
    • HXK2_HUMAN antibody
    • Muscle form hexokinase antibody
    see all

Images

  • All lanes : Anti-Hexokinase II antibody [EPR20839] (ab209847) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
    Lane 2 : HKDC1-knockdown HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 3 : Hexokinase I-knockdown HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 4 : Hexokinase II-knockdown HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 102 kDa
    Observed band size: 102 kDa


    Exposure time: 41 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Hexokinase II with ab209847 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining in HeLa cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human pancreatic adenocarcinoma tissue labeling Hexokinase II with ab209847 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining in human pancreatic adenocarcinoma (PMID: 26137268) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Hexokinase II with ab209847 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Hexokinase II was immunoprecipitated from 0.35 mg HeLa (humanepithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209847 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209847 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10 μg (Input).
    Lane 2: ab209847 IP in HeLa whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209847 in HeLa whole cell lysate.

    Exposure time: 30 seconds.

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling Hexokinase II with ab209847 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining in NIH/3T3 cells. 

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • All lanes : Anti-Hexokinase II antibody [EPR20839] (ab209847) at 1/1000 dilution

    Lane 1 : Rat testis lysate at 20 µg
    Lane 2 : Rat muscle lysate at 20 µg
    Lane 3 : SH-SY5Y (human neuroblastoma cell line from bone marrow), whole cell lysate at 20 µg
    Lane 4 : Human testis lysate at 20 µg
    Lane 5 : Mouse muscle lysate at 20 µg
    Lane 6 : Human colon lysate at 10 µg
    Lane 7 : Human fetal heart lysate at 10 µg
    Lane 8 : NIH/3T3 (mouse embryonic fibroblast cell line), whole cell lysate at 10 µg

    Secondary
    Lanes 1-5 & 8 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 6-7 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 102 kDa
    Observed band size: 102 kDa



     

    Exposure times: Lanes 1-3,6,7: 3 minutes; Lanes 4,5,8: 58 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Immunohistochemical analysis of paraffin-embedded human heart tissue labeling Hexokinase II with ab209847 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granularly cytoplasmic staining in human heart (PMID: 4058069; PMID: 26722360) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • All lanes : Anti-Hexokinase II antibody [EPR20839] (ab209847) at 1/10000 dilution

    Lane 1 : HKDC1 recombinant protein (aa16-458)
    Lane 2 : Hexokinase II recombinant protein (aa16-458)
    Lane 3 : Hexokinase I recombinant protein (aa16-458)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 102 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


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

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

References

This product has been referenced in:

  • Jiang Z  et al. Increased glycolysis correlates with elevated immune activity in tumor immune microenvironment. EBioMedicine 42:431-442 (2019). Read more (PubMed: 30935888) »
  • Lobato-Márquez D  et al. A requirement for septins and the autophagy receptor p62 in the proliferation of intracellular Shigella. Cytoskeleton (Hoboken) N/A:N/A (2018). Read more (PubMed: 29752866) »
See all 2 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab209847.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up