MED15 Antibody (monoclonal, 6F4)

SKU: orb548224

Description

Anti-MED15 Antibody (monoclonal, 6F4). Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

Images & Validation

• Tested Applications: FC, ICC, IF, WB
• Reactivity: Human
• Application Notes:
  Western blot, 0.1-0.5μg/ml Immunocytochemistry/Immunofluorescence, 2μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells. Add 0.2ml of distilled water will yield a concentration of 500μg/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG1
• Clone No.: 6F4
• Immunogen: E. coli-derived human MED15 recombinant protein (Position: M1-A285).
• Molecular Weight: 86 kDa

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
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Mediator of RNA polymerase II transcription subunit 15; Activator-recruited cofactor 105 kDa component; ARC105; CTG repeat protein 7a; Mediator complex subunit 15; Positive cofactor 2 glutamine; Q-rich-associated protein; PC2 glutamine; Q-rich-associated protein; TPA-inducible gene 1 protein; TIG-1; Trinucleotide repeat-containing gene 7 protein; MED15; ARC105; CTG7A; PCQAP; TIG1; TNRC7

Flow Cytometry analysis of U20S cells using anti-MED15 antibody. Overlay histogram showing U20S 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 mouse anti-MED15 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 MED15 using anti-MED15 antibody. MED15 was detected in immunocytochemical section of A431 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 2 µg/mL mouse anti-MED15 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.

Western blot analysis of MED15 using anti-MED15 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 50 ug of sample under reducing conditions. Lane 1: human K562 whole cell lysates, Lane 2: human U87 whole cell lysates, Lane 3: human CACO-2 whole cell lysates, Lane 4: human HL-60 whole cell lysates, Lane 5: human HepG2 whole cell lysates, Lane 6: human Hela whole cell lysates, Lane 7: human PC-3 whole cell lysates. 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 mouse anti-MED15 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 MED15 at approximately 86KD. The expected band size for MED15 is at 86KD.

Quick Database Links

UniProt

Q96RN5