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Anti-Hexokinase II antibody [EPR20839] ab209847 is a rabbit monoclonal antibody that is used in Hexokinase II western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with HK2 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR20839 is cited in over 170 publications


Images

Western blot - Anti-Hexokinase II antibody [EPR20839] (AB209847), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Hexokinase II antibody [EPR20839] (AB209847), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [EPR20839] (AB209847), expandable thumbnail
  • Immunoprecipitation - Anti-Hexokinase II antibody [EPR20839] (AB209847), expandable thumbnail
  • Western blot - Anti-Hexokinase II antibody [EPR20839] (AB209847), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Tested
Expected
Expected
Rat
Expected
Tested
Expected
Expected
Expected
Recombinant fragment - Human
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/30

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant fragment - Human

Dilution info

1/1000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/100

Notes

-

Species

Human

Dilution info

1/100

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/60

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

Not recommended for rat and mouse tissues.

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

2 products for Alternative Product

Target data

Function

Catalyzes the phosphorylation of hexose, such as D-glucose and D-fructose, to hexose 6-phosphate (D-glucose 6-phosphate and D-fructose 6-phosphate, respectively) (PubMed:23185017, PubMed:26985301, PubMed:29298880). Mediates the initial step of glycolysis by catalyzing phosphorylation of D-glucose to D-glucose 6-phosphate (PubMed:29298880). Plays a key role in maintaining the integrity of the outer mitochondrial membrane by preventing the release of apoptogenic molecules from the intermembrane space and subsequent apoptosis (PubMed:18350175).

Alternative names

Hexokinase-2, Hexokinase type II, Hexokinase-B, Muscle form hexokinase, HK II, HK2

Recommended products

Anti-Hexokinase II antibody [EPR20839] ab209847 is a rabbit monoclonal antibody that is used in Hexokinase II western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with HK2 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR20839 is cited in over 170 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR20839

Purification technique

Affinity purification Protein A

Specificity

Not recommended for rat and mouse tissues in IHC.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

  • Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    Lanes 1 - 4: Merged signal (red and green). Green - ab209847 observed at 105 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab209847 was shown to react with Anti-Hexokinase II in wild-type HCT 116 cells in western blot with loss of signal observed in HK2 knockout cell line Human HK2 (Hexokinase II) knockout HCT116 cell line ab273721 (HK2 knockout cell lysate Human HK2 (Hexokinase II) knockout HCT116 cell lysate ab275250). Wild-type and HK2 knockout HCT 116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab209847 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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.

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

    Lane 1: Wild-type HCT116 cell lysate at 20 µg

    Lane 2: HK2 knockout HCT116 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: SH-SY5Y cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 102 kDa

    Observed band size: 105 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    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.

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

  • Immunoprecipitation - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Immunoprecipitation - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab209847 in HeLa whole cell lysate.

    Exposure time: 30 seconds.

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

    All lanes: Immunoprecipitation - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    Predicted band size: 102 kDa

    Observed band size: 102 kDa

  • Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    Lanes 1 - 4: Merged signal (red and green). Green - ab209847 observed at 110 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab209847 was shown to react with Hexokinase II in wild-type HEK-293 cells in western blot with loss of signal observed in HK2 knockout sample. Wild-type and HK2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab209847 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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.

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

    Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

    Lane 2: HK2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

    Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 4: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 102 kDa

    Observed band size: 110 kDa

  • Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

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

    Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 10 µg

    Lane 2: HKDC1-knockdown HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Lane 3: Hexokinase I-knockdown HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Lane 4: Hexokinase II-knockdown HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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: 41s

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209847).

  • Immunocytochemistry/ Immunofluorescence - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    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.

    All lanes: Western blot - 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, 2, 3, 4, 5 and 8: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Lanes 6 - 7: Western blot - VeriBlot for IP Detection Reagent (HRP) (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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    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.

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

  • Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Western blot - Anti-Hexokinase II antibody [EPR20839] (ab209847)

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

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

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

    Lane 1: HKDC1 recombinant protein (aa16-458) at 0.01 µg

    Lane 2: Hexokinase II recombinant protein (aa16-458) at 0.01 µg

    Lane 3: Hexokinase I recombinant protein (aa16-458) at 0.01 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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

    Exposure time: 3min

  • Flow Cytometry (Intracellular) - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    Intracellular 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 (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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Dot Blot - Anti-Hexokinase II antibody [EPR20839] (ab209847), expandable thumbnail

    Dot Blot - Anti-Hexokinase II antibody [EPR20839] (ab209847)

    Dot blot analysis of Hexokinase Type III/HK3 using Anti-Hexokinase Type III/HK3 antibody [EPR29196-29] ab321996 at 1:1000 (0.525 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

    Lane1: His-tagged human HK3 fragment

    Lane2: His-tagged human HK2 fragment

    Lane3: His-tagged human HK1 fragment

    Lane4: His-tagged human HKDC1 fragment

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

    This antibody does not cross-react with human HK1, HK2 and HKDC1.

    All lanes: Dot Blot - Anti-Hexokinase Type III/HK3 antibody [EPR29196-29] (Anti-Hexokinase Type III/HK3 antibody [EPR29196-29] ab321996) at 1/1000 dilution

    Lane 1: His-tagged human HK3 fragment at 5 ng

    Lane 2: His-tagged human HK2 fragment at 5 ng

    Lane 3: His-tagged human HK1 fragment at 5 ng

    Lane 4: His-tagged human HKDC1 fragment at 5 ng

    Secondary

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

    Exposure time: 180s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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