ROC1/RBX1 Rabbit Polyclonal Antibody

SKU: orb334530

Description

Anti-ROC1/RBX1 Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Research Area

Cell Biology, Epigenetics & Chromatin, Protein Biochemistry

Images & Validation

• Tested Applications: FC, ICC, IF, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Human
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human ROC1, identical to the related mouse sequence.
• Target: E3 ubiquitin-protein ligase RBX1
• Molecular Weight: 15 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 antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3. *This antibody is supplied in a stabilized formulation. Compatibility with conjugation reactions depends on the chemistry of the conjugation method used. For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
• 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

E3 ubiquitin-protein ligase RBX1; 6.3.2.-; Protein ZYP; RING finger protein 75; RING-box protein 1; Rbx1; Regulator of cullins 1; E3 ubiquitin-protein ligase RBX1, N-terminally processed; RBX1; RNF75, ROC1

Flow Cytometry analysis of A431 cells using anti-ROC1 antibody. Overlay histogram showing A431 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-ROC1 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 ROC1 using anti-ROC1 antibody. ROC1 was detected in immunocytochemical section of Hela 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 rabbit anti-ROC1 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit 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 ROC1 using anti-ROC1 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: Rat Testis Tissue Lysate, Lane 2: Rat Brain Tissue Lysate, Lane 3: Mouse Brain Tissue Lysate, Lane 4: Mouse Spleen Tissue Lysate. 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-ROC1 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 ROC1 at approximately 15 kDa. The expected band size for ROC1 is at 15 kDa.

Quick Database Links

Gene Symbol

E3 ubiquitin-protein ligase RBX1

UniProt

P62877