RAD50 Antibody

SKU: orb1145739

Description

Anti-RAD50 Antibody. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

Images & Validation

• Tested Applications: ELISA, FC, ICC, IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Application Notes:
  Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human 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

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human RAD50 recombinant protein (Position: N1224-H1312).
• Molecular Weight: 154 kDa
• Purification: Immunogen affinity purified.

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

Ubiquitin-like modifier-activating enzyme ATG7; ATG12-activating enzyme E1 ATG7; Autophagy-related protein 7; APG7-like; hAGP7; Ubiquitin-activating enzyme E1-like protein; ATG7; APG7L

Flow Cytometry analysis of A549 cells using anti-RAD50 antibody. Overlay histogram showing A549 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-RAD50 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of RAD50 using anti-RAD50 antibody. RAD50 was detected in an immunocytochemical section of A549 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 rabbit anti-RAD50 Antibody overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 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 RAD50 using anti-RAD50 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 Jurkat whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human K562 whole cell lysates, Lane 5: human A549 whole cell lysates, Lane 6: human Caco-2 whole cell lysates, Lane 7: human Daudi whole cell lysates, Lane 8: human SH-SY5Y 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-RAD50 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 RAD50 at approximately 154 kDa. The expected band size for RAD50 is at 154 kDa.

Quick Database Links

UniProt

Q92878