Name: Anti-Hexokinase II antibody [EPR20839]
SKU: ab209847
Rating: 5 (2 reviews)

Anti-Hexokinase II antibody [EPR20839] is a rabbit recombinant monoclonal antibody that is used to detect Hexokinase II in Flow cytometry (Intra), ICC/IF, IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.

- Cited in over 105 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with HK2 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Images

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.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow 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	Dilution info	Notes
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	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
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	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -
Species Human	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/60	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
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	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Associated Products

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2 products for Alternative Product

Target data

See full target information HK2

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

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.
Clone number: EPR20839
Purification technique: Affinity purification Protein A
Specificity: Not recommended for rat and mouse tissues in IHC.
Concentration

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

Product Specifications

Anti-Hexokinase II antibody [EPR20839] (ab209847) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P, IP, WB in human, mouse, rat samples.
Anti-Hexokinase II antibody [EPR20839] (ab209847) specifically detects Hexokinase II (UniProt ID: P52789; Molecular weight: 102kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation

Abcam's high quality manufacturing and validation processes ensure Anti-Hexokinase II antibody [EPR20839] (ab209847) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-Hexokinase II antibody [EPR20839] (ab209847) has been confirmed by testing in knockout samples.
Anti-Hexokinase II antibody [EPR20839] (ab209847) has been cited over 110 times in peer reviewed journals and is trusted by the scientific community.

Related Products
Conjugation-ready, carrier free format available for antibody clone EPR20839 - ab228819.
Antibody clone EPR20839 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 647, HRP (ab237314, ab237315).

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)
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 2: Western blot - Human HK2 (Hexokinase II) knockout HCT116 cell line (Human HK2 (Hexokinase II) knockout HCT116 cell line ab273721)
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)
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)
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)
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)
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 2: Western blot - Human HK2 (Hexokinase II) knockout HEK-293 cell line (Human HK2 (Hexokinase II) knockout HEK-293 cell line ab269485)
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)
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
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)
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)
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)
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)
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)
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