PP2A-alpha/PPP2CA/PP2A Mouse Monoclonal Antibody

SKU: orb654389

Description

Anti-PP2A-alpha/PPP2CA Antibody (monoclonal, 3B6). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat.

Research Area

Cell Biology, Disease Biomarkers, Neuroscience, Protein Biochemistry, Signal Transduction

Images & Validation

• Tested Applications: FC, ICC, IF, IHC, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml, Human, Monkey, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat Immunocytochemistry/Immunofluorescence, 4μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Human
• Reactivity: Human, Monkey, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG2a
• Clone No.: 3B6
• Immunogen: E.coli-derived human PP2A-alpha recombinant protein (Position: M1-L309). Human PP2A-alpha shares 100% amino acid (aa) sequence identity with both mouse and rat PP2A-alpha.
• Target: Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform
• Molecular Weight: 36 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
• Concentration: 500 µg/ml
• Disclaimer: For research use only

Alternative Names

PP2A alpha; PP2Ac; PP2CA; PPP2CA; Replication protein C; RP C

Flow Cytometry analysis of U937 cells using anti-PP2A-alpha/PPP2CA antibody. Overlay histogram showing U937 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-PP2A-alpha/PPP2CA 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of PP2A-alpha/PPP2CA using anti-PP2A-alpha/PPP2CA antibody. PP2A-alpha/PPP2CA was detected in immunocytochemical section of HELA cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4 µg/mL mouse anti-PP2A-alpha/PPP2CA Antibody 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.

IHC analysis of PP2A-alpha/PPP2CA using anti-PP2A-alpha/PPP2CA antibody. PP2A-alpha/PPP2CA 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 1 µg/ml mouse anti-PP2A-alpha/PPP2CA Antibody 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) with DAB as the chromogen.

IHC analysis of PP2A-alpha/PPP2CA using anti-PP2A-alpha/PPP2CA antibody. PP2A-alpha/PPP2CA was detected in paraffin-embedded section of human rectal 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 1 µg/ml mouse anti-PP2A-alpha/PPP2CA Antibody 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) with DAB as the chromogen.

IHC analysis of PP2A-alpha/PPP2CA using anti-PP2A-alpha/PPP2CA antibody. PP2A-alpha/PPP2CA was detected in paraffin-embedded section of rat brain 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 1 µg/ml mouse anti-PP2A-alpha/PPP2CA Antibody 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) with DAB as the chromogen.

Western blot analysis of PP2A-alpha/PPP2CA using anti-PP2A-alpha/PPP2CA antibody. 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 50 ug of sample under reducing conditions. Lane 1: human HELA whole cell lysates, Lane 2: human HEPG2 whole cell lysates, Lane 3: monkey COS-7 whole cell lysates, Lane 4: rat PC-12 whole cell lysates, Lane 5: mouse NIH/3T3 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 mouse anti-PP2A-alpha/PPP2CA 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. A specific band was detected for PP2A-alpha/PPP2CA at approximately 36 KD. The expected band size for PP2A-alpha/PPP2CA is at 36 KD.

Quick Database Links

Gene Symbol

Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform

UniProt

P67775