Anti-SAPS3/PPP6R3 Antibody

• Catalog Number: orb1972544
• Category: Antibodies
• Description: Anti-SAPS3/PPP6R3 Antibody. Tested in WB, ICC/IF, Flow Cytometry, ELISA applications. This antibody reacts with Human, Monkey, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, ICC, IF, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Immunogen: E.coli-derived human SAPS3/PPP6R3 recombinant protein (Position: R279-R848).
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 144 kDa
• UniProt ID: Q5H9R7
• 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.
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human, Monkey, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human ELISA, 0.1-0.5 μg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of Hela cells using anti-SAPS3/PPP6R3 antibody. Overlay histogram showing Hela 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-SAPS3/PPP6R3 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 (Red line) was also used as a control.

IF analysis of PPP6R3 using anti-PPP6R3 antibody and anti-Beta Tubulin antibody. PPP6R3 was detected in immunocytochemical section of MCF-7 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-PPP6R3 Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and DyLight®550 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of SAPS3/PPP6R3 using anti-SAPS3/PPP6R3 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 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human U-87MG whole cell lysates, Lane 4: monkey COS-7 whole cell lysates, Lane 5: rat NRK whole cell lysates, Lane 6: 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 rabbit anti-SAPS3/PPP6R3 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 Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for SAPS3/PPP6R3 at approximately 144 kDa. The expected band size for SAPS3/PPP6R3 is at 95 kDa.