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Product Info Summary
| SKU: | A01023-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-ROCK2 Antibody Picoband®
SKU/Catalog Number
A01023-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ROCK2 Antibody Picoband® catalog # A01023-1. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-ROCK2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01023-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ROCK2 recombinant protein (Position: E652-D909).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A01023-1 is reactive to ROCK2 in Human, Mouse, Rat
Observed Molecular Weight
161 kDa
Calculated molecular weight
160.9 kDa
Background of ROCK2
Rho-associated kinase (ROCK), including the ROCK-I and ROCK-II isoforms, is a protein kinase involved in signaling from Rho to actin cytoskeleton. Serine/threonine kinase ROCK II/Rho kinase, which is an isozyme of ROCK I, is one of the targets for the small GTPase Rho. ROCK II regulates the formation of actin stress fibers and focal adhesions, cytokinesis, smooth muscle contraction, and the activation of c-fos serum response element. Sequencing analysis has shown that human ROCK II contains 1388 amino acid residues with a calculated molecular mass of approximately 161 kDa. Fluorescence in situ hybridization analysis showed that the human ROCK II gene is located on chromosome 2p24. Thumkeo et al. concluded that ROCK-II is essential in inhibiting blood coagulation and maintaining blood flow in the endothelium-free labyrinth layer and that loss of ROCK-II leads to thrombus formation, placental dysfunction, intrauterine growth retardation, and fetal death.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A01023-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
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, -
Positive Control
WB: human HELA whole cell, human K562 whole cell, human Jurkat whole cell, human HT1080 whole cell, human U-87MG whole cell, human CACO-2 whole cell, rat brain tissue, rat PC-12 whole cell, mouse brain tissue, mouse lung tissue, mouse NIH/3T3 whole cell
ICC/IF: Hep cell
FCM: A549 cell
Validation Images & Assay Conditions
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Western blot analysis of ROCK2 using anti-ROCK2 antibody (A01023-1).
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 50ug 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 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 rabbit anti-ROCK2 antigen affinity purified polyclonal antibody (Catalog # A01023-1) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ROCK2 at approximately 161KD. The expected band size for ROCK2 is at 161KD.
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Flow Cytometry analysis of A549 cells using anti-ROCK2 antibody (A01023-1).
Overlay histogram showing A549 cells stained with A01023-1 (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 (A01023-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IF analysis of ROCK2 using anti-ROCK2 antibody (A01023-1).
ROCK2 was detected in immunocytochemical section of Hep cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-ROCK2 Antibody (A01023-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
Specific Publications For Anti-ROCK2 Antibody Picoband® (A01023-1)
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