Anti-AMPK beta 2 PRKAB2 Antibody (monoclonal, 6G1)

• Catalog Number: orb443138
• Category: Antibodies
• Description: Anti-AMPK beta 2 PRKAB2 Antibody (monoclonal, 6G1). Tested in Flow Cytometry, IHC, ICC, WB applications. This antibody reacts with Human.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 6G1
• Tested applications: FC, ICC, IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG2b
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human AMPK beta 2, different from the related mouse sequence by three amino acids, and from the related rat sequence by two amino acids.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 34 kDa
• UniProt ID: O43741
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: 5'-AMP-activated protein kinase subunit beta-2; AM
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Frozen Section), 0.5-1μg/ml Immunocytochemistry, 0.5-1μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of PC-3 cells using anti-AMPK beta 2 antibody. Overlay histogram showing PC-3 cells (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 mouse anti-AMPK beta 2 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Western blot analysis of AMPK beta 2 using anti-AMPK beta 2 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysate, Lane 2: human placenta tissue lysate, Lane 3: human 293T whole cell lysate, Lane 4: human A549 whole cell lysate, Lane 5: human A375 whole cell lysate, Lane 6: human A431 whole cell lysate, Lane 7: human U20S whole cell lysate, Lane 8: human K562 whole cell lysate. 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-AMPK beta 2 antigen affinity purified monoclonal antibody 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 with Tanon 5200 system.