POLR2D Rabbit Polyclonal Antibody

SKU: orb1743909

Description

Anti-POLR2D Antibody. Tested in ELISA, IF, ICC, WB applications. This antibody reacts with Human.

Research Area

Epigenetics & Chromatin, Molecular Biology

Images & Validation

• Tested Applications: ELISA, WB
• Dilution Range: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat ELISA, 0.1-0.5 μg/ml, -
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human POLR2D recombinant protein (Position: D7-D124).
• Target: DNA-directed RNA polymerase II subunit RPB4
• Molecular Weight: 18 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, 0.2 mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Mediator of RNA polymerase II transcription subunit 9; Mediator complex subunit 9; MED9; MED25

IF analysis of POLR2D using anti-POLR2D antibody and anti-Beta Tubulin antibody. POLR2D 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-POLR2D Antibody and mouse anti-Beta Tubulin antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG and DyLight®488 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 POLR2D using anti-POLR2D 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 MCF-7 whole cell lysates, Lane 2: human 293T 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-POLR2D 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 POLR2D at approximately 18 kDa. The expected band size for POLR2D is at 16-20 kDa.

Quick Database Links

Gene Symbol

DNA-directed RNA polymerase II subunit RPB4

UniProt

O15514