Anti-ROCK2 Antibody Picoband™

Product Name

Anti-ROCK2 Antibody Picoband™

View all ROCK2 Antibodies

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.

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).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A01023-1 is reactive to ROCK2 in Human, Mouse, Rat

Applications

A01023-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

161 kDa

Calculated molecular weight

160.9kDa

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. Flow Cytometry analysis of A549 cells using anti-ROCK2 antibody (A01023-1).
Overlay histogram showing A549 cells stained with A01023-1 (Blue line).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 (Red line) was also used as a control.

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Figure 3. 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.

Gene/Protein Information For ROCK2 (Source: Uniprot.org, NCBI)

Alternative Names

coiled-coil-containing protein kinase 2; EC 2.7.11; EC 2.7.11.1; KIAA0619; p164 ROCK-2; Rho kinase 2; rho-associated protein kinase 2; Rho-associated, coiled-coil containing protein kinase 2; ROCK2; ROCK-II; ROKalpha ROCK2 ROCK-II Rho associated coiled-coil containing protein kinase 2 rho-associated protein kinase 2|p164 ROCK-2|rho-associated, coiled-coil-containing protein kinase II

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