Phospho-POLR2A (S5) Rabbit Monoclonal Antibody

• Catalog Number: orb548621
• Category: Antibodies
• Description: Phospho-POLR2A (S5) Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: ABCE-16
• Tested applications: FC, ICC, IF, IHC, IP, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from human POLR2A
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 217176 MW
• UniProt ID: P24928
• 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: DNA-directed RNA polymerase II subunit RPB1;RNA po
• Note: For research use only
• Application notes: WB 1:500-1:2000IHC 1:50-1:200ICC/IF 1:50-1:200IP 1:20FC 1:40
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Phospho-POLR2A (S5) expression in (1) HeLa cell lysate treated with Lambda Phosphatase (orb548621) ; (2) HeLa cell lysate. 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 50ug of sample under reducing conditions. 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 rabbit anti-POLR2A monoclonal antibody (Catalog # orb548621) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system.

IHC analysis of Phospho-POLR2A (S5) using anti-Phospho-POLR2A (S5) antibody (orb548621) on human spleen. Phospho-POLR2A (S5) was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-Phospho-POLR2A (S5) Antibody (orb548621) overnight at 4°C. Biotinylated goat anti Rabbit IgG IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.

IF analysis of immunocytochemical section of HeLa cells using anti-Phospho-POLR2A (S5) antibody (orb548621) Phospho-POLR2A (S5) was detected in immunocytochemical section. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL rabbit anti-Phospho-POLR2A (S5) Antibody (orb548621) overnight at 4 &deg.