WB
Western blot analysis of Hexokinase II on different lysates with Rabbit anti-Hexokinase II antibody at 1/1000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: HCT 116 cell lysate (20 µg/Lane), Lane 3: NIH/3T3 cell lysate (20 µg/Lane), Lane 4: PC-12 cell lysate (20 µg/Lane), Lane 5: COS-1 cell lysate (20 µg/Lane), Lane 6: Mouse skeletal muscle tissue lysate (40 µg/Lane), Lane 7: Mouse testis tissue lysate (40 µg/Lane), Lane 8: Rat skeletal muscle tissue lysate (40 µg/Lane), Lane 9: Rat testis tissue lysate (40 µg/Lane), Exposure time: 8 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Hexokinase II with Rabbit anti-Hexokinase II antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hexokinase II antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L ( 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.ICC/IF
Immunocytochemistry analysis of PC-12 cells labeling Hexokinase II with Rabbit anti-Hexokinase II antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hexokinase II antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Hexokinase II antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-Hexokinase II antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-Hexokinase II antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibodyat 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IP
Hexokinase II was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-Hexokinase II antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Hexokinase II antibody at 1/2000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-Hexokinase II antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Hexokinase II antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 6 seconds.FC
Flow cytometric analysis of NIH/3T3 cells labeling Hexokinase II. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).FC
Flow cytometric analysis of PC-12 cells labeling Hexokinase II. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Hexokinase II Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000 IHC:1:50 ICC/IF:1:50-1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | HK2 |
|---|---|
| Gene Synonyms | HKII HXK2 |
| Gene Full Name | hexokinase 2 |
| Gene Summary | Hexokinases phosphorylate glucose to produce glucose-6-phosphate, the first step in most glucose metabolism pathways. This gene encodes hexokinase 2, the predominant form found in skeletal muscle. It localizes to the outer membrane of mitochondria. Expression of this gene is insulin-responsive, and studies in rat suggest that it is involved in the increased rate of glycolysis seen in rapidly growing cancer cells. [provided by RefSeq, Apr 2009] |
| 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 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 |
| Molecular Weight(MW) | 102kDa(Observed band size:102kDa) |
| Cellular Localization | Mitochondrion outer membrane, Cytoplasm, cytosol. |
WB
Western blot analysis of Hexokinase II on different lysates with Rabbit anti-Hexokinase II antibody at 1/1000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: HCT 116 cell lysate (20 µg/Lane), Lane 3: NIH/3T3 cell lysate (20 µg/Lane), Lane 4: PC-12 cell lysate (20 µg/Lane), Lane 5: COS-1 cell lysate (20 µg/Lane), Lane 6: Mouse skeletal muscle tissue lysate (40 µg/Lane), Lane 7: Mouse testis tissue lysate (40 µg/Lane), Lane 8: Rat skeletal muscle tissue lysate (40 µg/Lane), Lane 9: Rat testis tissue lysate (40 µg/Lane), Exposure time: 8 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Hexokinase II with Rabbit anti-Hexokinase II antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hexokinase II antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L ( 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
ICC/IF
Immunocytochemistry analysis of PC-12 cells labeling Hexokinase II with Rabbit anti-Hexokinase II antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hexokinase II antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Hexokinase II antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-Hexokinase II antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-Hexokinase II antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibodyat 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IP
Hexokinase II was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-Hexokinase II antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Hexokinase II antibody at 1/2000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-Hexokinase II antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Hexokinase II antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 6 seconds.
FC
Flow cytometric analysis of NIH/3T3 cells labeling Hexokinase II. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
FC
Flow cytometric analysis of PC-12 cells labeling Hexokinase II. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000 IHC:1:50 ICC/IF:1:50-1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
