Anti-Hexokinase II/HK2 Antibody Picoband®

The influences of circPOSTN silencing on proliferation, apoptosis and aerobic glycolysis of glioma cells. a – l LN229 and U251 cells were transfected with si-circPOSTN or si-NC. a The interference efficiency of si-circPOSTN was analyzed with RT-qPCR assay in LN229 and U251 cells. b , c Effect of circPOSTN silencing on the cell viability of LN229 and U251 cells was assessed with MTT assay. d The apoptosis rate was computed with flow cytometry assay in transfected LN229 and U251 cells. e The western blot assay showed the expression levels of Bcl-2 and Bax in LN229 and U251 cells. f The caspase-3 activity was measured with a caspase-3 assay kit. g – i The concentration of glucose and lactate in the culture medium, as well as ATP production level were measured with a series of kits, respectively. j The protein expression levels of HK2 and LDHA were determined with western blot assay in transfected LN229 and U251 cells. k – l LDHA enzyme activity and ROS accumulation were evaluated in LN229 and U251 cells post-transfection with lactate dehydrogenase activity detection kit and reactive oxygen species assay kit, respectively. * P < 0.05
Index in PubMed under a CC BY license. PMID: 32774168

CircPOSTN silencing inhibited aerobic glycolysis of glioma cells via regulating miR-361-5p. a – l LN229 and U251 cells were transfected with si-NC, si-circPOSTN, si-circPOSTN + anti-miR-NC, or si-circPOSTN + anti-miR-361-5p. a – f The concentration of glucose and lactate, as well as cellular ATP level were detected with different kits. g , h The protein expression levels of HK2 and LDHA in LN229 and U251 cells were measured with western blot assay. i – l The enzyme activity of LDHA and ROS level were measured in transfected LN229 and U251 cells. * P < 0.05
Index in PubMed under a CC BY license. PMID: 32774168

TPX2 regulated proliferation, apoptosis, and aerobic glycolysis in glioma cells. a – l LN229 and U251 cells were introduced with si-NC or si-TPX2. a The transfection efficiency of si-TPX2 was checked with RT-qPCR assay in LN229 and U251 cells. b , c The cell viability of LN229 and U251 cells was determined with MTT assay. d The apoptosis rate of transfected LN229 and U251 cells was represented by flow cytometry assay. e The western blot assay was used to assay the expression levels of Bcl-2 and Bax in LN229 and U251 cells. f The activity of caspase-3 was detected with a caspase-3 assay kit. g – i The glucose, lactate, and ATP production levels were shown. j The protein expression levels of HK2 and LDHA were estimated by western blot assay in LN229 and U251 cells. k , l LDHA enzyme activity and ROS content were evaluated in LN229 and U251 cells post-transfection. * P < 0.05
Index in PubMed under a CC BY license. PMID: 32774168

Western blot analysis of Hexokinase 2/HK2 using anti-Hexokinase 2/HK2 antibody (A01389).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela- WT whole cell lysates,
Lane 2: human Hela-HK2 KO whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hexokinase 2/HK2 antigen affinity purified polyclonal antibody (A01389) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Hexokinase 2/HK2 at approximately 102 kDa. The expected band size for Hexokinase 2/HK2 is at 102 kDa.

Western blot analysis of Hexokinase II using anti-Hexokinase II antibody (A01389).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: rat skeletal muscle tissue lysates,
Lane 5: rat heart tissue lysates,
Lane 6: mouse skeletal muscle tissue lysates,
Lane 7: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hexokinase II antigen affinity purified polyclonal antibody (A01389) 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-rabbit IgG-HRP secondary antibody (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Hexokinase II at approximately 102 kDa. The expected band size for Hexokinase II is at 102 kDa.

IHC analysis of Hexokinase II using anti-Hexokinase II antibody (A01389).
Hexokinase II was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Hexokinase II Antibody (A01389) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Hexokinase II using anti-Hexokinase II antibody (A01389).
Hexokinase II was detected in a paraffin-embedded section of mouse skeletal muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Hexokinase II Antibody (A01389) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Hexokinase II using anti-Hexokinase II antibody (A01389).
Hexokinase II was detected in a paraffin-embedded section of rat skeletal muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Hexokinase II Antibody (A01389) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Flow Cytometry analysis of HepG2 cells using anti-Hexokinase II antibody (A01389).
Overlay histogram showing HepG2 cells stained with A01389 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Hexokinase II Antibody (A01389, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.