|Reactivity||Human, Mouse, Rat|
|Product Name||Anti-ROCK2 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Rho-associated protein kinase 2(ROCK2) detection. Tested with WB, IHC-P in Human;Mouse;Rat.|
|Cite This Product||Anti-ROCK2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1242)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Immunogen||A synthetic peptide corresponding to a sequence at the N-terminus of human ROCK2(35-51aa RDPRSPINVESLLDGLN), identical to the related rat and mouse sequences.|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Anti-ROCK2 antibody, PA1242, Western blotting
Lane 1: MCF-7 Cell Lysate
Lane 2: HELA Cell Lysate
Lane 3: SMMC Cell Lysate
Lane 4: MM453 Cell Lysate
Anti-ROCK2 antibody, PA1242, IHC(P)
IHC(P): Human Rectal Cancer Tissue
Figure 3. Western blot analysis of ROCK2 using anti-ROCK2 antibody (PA1242).
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: rat brain tissue lysate.
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 # PA1242) at 0.5 Î¼g/mL overnight at 4Â°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 1:10000 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.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Rho-associated protein kinase 2|
|Alternative Names||Rho-associated protein kinase 2;18.104.22.168;Rho kinase 2;Rho-associated, coiled-coil-containing protein kinase 2;Rho-associated, coiled-coil-containing protein kinase II;ROCK-II;p164 ROCK-2;ROCK2;KIAA0619;|
|Subcellular Localization||Cytoplasm. Cell membrane ; Peripheral membrane protein . Nucleus. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Cytoplasmic, and associated with actin microfilaments and the plasma membrane. .|
|Molecular Weight||160900 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of ADD1, BRCA2, CNN1, EZR, DPYSL2, EP300, MSN, MYL9/MLC2, NPM1, RDX, PPP1R12A and VIM. Phosphorylates SORL1 and IRF4. Acts as a negative regulator of VEGF-induced angiogenic endothelial cell activation. Positively regulates the activation of p42/MAPK1-p44/MAPK3 and of p90RSK/RPS6KA1 during myogenic differentiation. Plays an important role in the timely initiation of centrosome duplication. Inhibits keratinocyte terminal differentiation. May regulate closure of the eyelids and ventral body wall through organization of actomyosin bundles. Plays a critical role in the regulation of spine and synaptic properties in the hippocampus. Plays an important role in generating the circadian rhythm of the aortic myofilament Ca(2+) sensitivity and vascular contractility by modulating the myosin light chain phosphorylation. .|
|Research Areas||Human, Mouse, Rat
*You can search these to find other products in these research areas.
|Background||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.(2003) 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.|
Other Recommended Resources
Here are featured tools and databases that you might find useful.
Download troubleshooting handbooks for IHC, Western blot and ELISA for FREE.Download Free PDFs Now
Guaranteed product quality
We promise all of our products perform as described in datasheets.
Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at email@example.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact firstname.lastname@example.org
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.