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Catalog Number | orb654274 |
---|---|
Category | Antibodies |
Description | Hexokinase 1/HK1 Antibody (monoclonal, 4B7) |
Species/Host | Mouse |
Clonality | Monoclonal |
Clone Number | 4B7 |
Tested applications | FC, ICC, IF, IHC, IHC-Fr, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Mouse IgG2a |
Immunogen | E.coli-derived human Hexokinase 1/HK1 recombinant protein (Position: D17-R323). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human Immunofluorescence, 2μg/ml, Human Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 120 kDa |
UniProt ID | P19367 |
Storage | Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles. |
Alternative names | Hexokinase-1; Brain form hexokinase; Hexokinase ty Read more... |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HeLa whole cell lysates; Lane 2: human HEK293 whole cell lysates; Lane 3: human U-87MG whole cell lysates; Lane 4: human K562 whole cell lysates; Lane 5: human A549 whole cell lysates; Lane 6: rat PC-12 whole cell lysates; Lane 7: mouse NIH/3T3 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Hexokinase 1/HK1 antigen affinity purified monoclonal antibody (Catalog # orb654274) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for Hexokinase 1/HK1 at approximately 120KD. The expected band size for Hexokinase 1/HK1 is at 120KD.
IHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in paraffin-embedded section of human testis cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
IF analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IF analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of PC-3 cells using anti-Hexokinase 1/HK1 antibody (orb654274). Overlay histogram showing PC-3 cells stained with orb654274 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Hexokinase 1/HK1 Antibody (orb654274, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
IHC analysis of Hexokinase 1/HK1 using anti-Hexokinase 1/HK1 antibody (orb654274). Hexokinase 1/HK1 was detected in a frozen section of human placenta tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hexokinase 1/HK1 Antibody (orb654274) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.
FC, ICC, IF, IHC, IHC-Fr, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
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