PDE6 beta/PDE6B Mouse Monoclonal Antibody

SKU: orb865587

Description

Anti-PDE6 beta/PDE6B Antibody (monoclonal, 8I2F10). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Research Area

Cancer Biology, Cell Biology, Disease Biomarkers, Metabolism Research, Neuroscience, Signal Transduction

Images & Validation

• Tested Applications: FC, ICC, IF, IHC, WB
• Dilution Range: Western blot, 0.25-0.5 μg/ml, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x^6 cells, Human
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG2b
• Clone No.: 8I2F10
• Immunogen: E.coli-derived human PDE6 beta/PDE6B recombinant protein (Position: K25-Q237).
• Target: Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta
• Molecular Weight: 98 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
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

CSNB3; GMP PDE beta; PDE6B; PDEB; rd1; RP40

Flow Cytometry analysis of U20S cells using anti-PDE6 beta/PDE6B antibody. Overlay histogram showing U20S cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with mouse anti-PDE6 beta/PDE6B Antibody (1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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.

IF analysis of PDE6 beta/PDE6B using anti-PDE6 beta/PDE6B antibody. PDE6 beta/PDE6B was detected in an immunocytochemical section of SH-SY5Y 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 5 µg/mL mouse anti-PDE6 beta/PDE6B 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 PDE6 beta/PDE6B using anti-PDE6 beta/PDE6B antibody. PDE6 beta/PDE6B was detected in a paraffin-embedded section of rat eye ball 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 mouse anti-PDE6 beta/PDE6B 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 PDE6 beta/PDE6B using anti-PDE6 beta/PDE6B 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: rat eye tissue lysates, Lane 2: mouse eye 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 mouse anti-PDE6 beta/PDE6B 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 PDE6 beta/PDE6B at approximately 98 kDa. The expected band size for PDE6 beta/PDE6B is at 98 kDa.

Quick Database Links

Gene Symbol

Rod cGMP-specific 3',5'-cyclic phosphodiesterase subunit beta

UniProt

P35913