Anti-PPP2R5D Antibody

• Catalog Number: orb1728167
• Category: Antibodies
• Description: Anti-PPP2R5D Antibody
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Purification: Immunogen affinity purified.
• Immunogen: E.coli-derived human PPP2R5D recombinant protein (Position: R91-L602).
• UniProt ID: Q14738
• MW: 74 kDa
• Tested applications: ELISA, FC, ICC, IF, IHC, IP, WB
• Application notes: Western blot, 0.1-0.25 μg/ml, Human, Mouse, Rat Immunohistochemistry, 2-5 μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Immunoprecipitation, 0.5-2 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x1x106 cells, Human ELISA, 0.1-0.5 μg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Cross Reactivity: No cross-reactivity with other proteins.
• Antibody Type: Primary Antibody
• 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: Semaphorin-3C; Semaphorin-E; Sema E; SEMA3C; SEMAE
• Note: For research use only

Flow Cytometry analysis of 293T cells using anti-PPP2R5D antibody. Overlay histogram showing 293T 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 rabbit anti-PPP2R5D Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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 PPP2R5D using anti-PPP2R5D antibody and anti-Tubulin Alpha antibody. PPP2R5D was detected in immunocytochemical section of U2OS cell. 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 5 µg/mL rabbit anti-PPP2R5D Antibody and mouse anti-Tubulin Alpha antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and Cy3 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 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 PPP2R5D using anti-PPP2R5D antibody. PPP2R5D was detected in a paraffin-embedded section of human stomach 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 2 µg/ml rabbit anti-PPP2R5D Antibody 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 with DAB as the chromogen.

Immunoprecipitating PPP2R5D in HepG2 whole cell lysate. Western blot analysis of PPP2R5D using anti-PPP2R5D antibody; Lane 1: HepG2 whole cell lysates (30 ug); Lane 2: Rabbit control IgG instead of anti-PPP2R5D antibody in HepG2 whole cell lysate; Lane 3: anti-PPP2R5D antibody (2 µg) + HepG2 whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PPP2R5D antigen affinity purified polyclonal antibody at a dilution of 0.5 µg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for PPP2R5D at approximately 74 kDa. The expected band size for PPP2R5D is at 70 kDa.

Western blot analysis of PPP2R5D using anti-PPP2R5D antibody. Electrophoresis was performed on a 10% 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 HepG2 whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: rat brain tissue lysates, Lane 5: mouse brain 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-PPP2R5D antigen affinity purified polyclonal 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-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 with Tanon 5200 system. A specific band was detected for PPP2R5D at approximately 74 kDa. The expected band size for PPP2R5D is at 70 kDa.