Anti-ROCK2 Antibody

• Catalog Number: orb738394
• Category: Antibodies
• Description: Anti-ROCK2 Antibody. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human ROCK2 recombinant protein (Position: E652-D909).
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 161 kDa
• UniProt ID: O75116
• 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: Complement receptor type 1; C3b/C4b receptor; CD35
• Note: For research use only
• Application notes: Western blot, 0.1-0.25μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5μg/ml, Human, Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of A549 cells using anti-ROCK2 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-ROCK2 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 ROCK2 using anti-ROCK2 antibody. ROCK2 was detected in immunocytochemical section of Hep 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-ROCK2 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 ROCK2 using anti-ROCK2 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 HELA whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human HT1080 whole cell lysates, Lane 5: human U-87MG whole cell lysates, Lane 6: human CACO-2 whole cell lysates, Lane 7: rat brain tissue lysates, Lane 8: rat PC-12 whole cell lysates, Lane 9: mouse brain tissue lysates, Lane 10: mouse lung tissue lysates, Lane 11: mouse NIH/3T3 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-ROCK2 antigen affinity purified polyclonal antibody at 0.25 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 ROCK2 at approximately 161 KD. The expected band size for ROCK2 is at 161 KD.