GeneTex
United States (US)

ROCK1 antibody

ROCK1-antibody_WB_GTX10549-1_19011506_284.jpg

Cat No. GTX10549

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB
Reactivity Human, Mouse, Dog
APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Dilution
WB Assay dependent
Not tested in other applications.

Calculated MW

158 kDa. ( Note )

Positive Control

MDCK cells treated with staurosporine
PROPERTIES

Form

Liquid

Buffer

PBS pH 7.3 (without Mg2+ and Ca2+) containing 50% glycerol, 1.0mg/ml BSA (IgG, protease free). and 0.05% sodium azide.

Storage

Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.

Antigen Species

Human

Immunogen

Synthetic peptide (Human).

Purification

Immunogen affinity purified
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a spanning peptide corresponding to the cleavage-site to remove antibody that is reactive with non-cleavage-site specific ROCK1. T

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

Rho Associated Coiled-Coil Containing Protein Kinase 1,P160Rock,Rock-I,Rock1

Cellular Localization

Cytoplasmic

Background

ROCK1, a 160 kDa small-GTP-effector protein, is a serine/threonine kinase involved in the induction of focal adhesions and stress fibers, and can play a major role in vascular smooth muscle contraction, malignant cell transformation, tumor invasion and metastasis. ROCK1 is identified as a substrate for caspases and is cleaved by caspase 3 at a conserved DETD1113/G sequence which removes its carboxy-terminal inhibitory domain, resulting in a constitutively active 130 kDa ROCK1 kinase. Proteolytic activation of ROCK1 by caspase is critical for its role in regulating the actin cytoskeleton and Myosin Light Chain (MLC) phosphorylation. Increased phosphorylation of MLC is important for membrane blebbing, a cellular event that occurs at the onset of apoptosis.

Database

Research Area

DATA IMAGES
ROCK1-antibody_WB_GTX10549-1_19011506_284.jpg

GTX10549 WB Image

Lysates prepared from MDCK cells left untreated (1, 3) or treated with staurosporine (2) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ROCK1 (GTX10549) cleavage-site specific antibody (1, 2) or a ROCK1 pan antibody (3) in a 3% BSA-TBST buffer for two hours at room temperature. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected. The data show that ROCK1 [1113/1114] CSSA detected the 130 kDa cleaved form of ROCK but not the full length. This antibody did not detect the 130 kDa cleaved form of ROCK upon inhibition of caspase activity with the pan caspase inhibitor Ac-VAD-CHO (data not shown).