Product nameAnti-ROCK2 antibody
See all ROCK2 primary antibodies
DescriptionRabbit polyclonal to ROCK2
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow
- This antobody gave a positive signal in the followign Lysates: Mouse Liver Tissue SK N BE Whole Cell Human Liver Tissue HeLa Whole Cell - Hydroxyurea Treated (48hr, 2uM) Y79 Whole Cell Rat Liver Tissue HeLa Whole Cell - Staurosporine Treated (24hr, 500nM) Raji Whole Cell Jurkat Whole Cell - Staurosporine Treated (24hr, 500nM) HeLa Whole Cell Lysate - Bleomycin Treated (20U/ml)
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab66320 in the following tested applications.
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 171 kDa (predicted molecular weight: 161 kDa).|
FunctionRegulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction.
Sequence similaritiesBelongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 PH domain.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain.
Contains 1 REM (Hr1) repeat.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationCytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane.
- Information by UniProt
- coiled-coil-containing protein kinase 2 antibody
- KIAA0619 antibody
- p164 ROCK 2 antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ROCK2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Rat liver tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab66320 observed at 171 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab66320 was shown to recognize ROCK2 when ROCK2 knockout samples were used, along with additional cross-reactive bands. Wild-type and ROCK2 knockout samples were subjected to SDS-PAGE. ab66320 at a concentration of 2 µg/ml and ab18058 (loading control to Vinculin) at a dilution of 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-ROCK2 antibody (ab66320) at 1 µg/ml
Lane 1 : Liver (Mouse) Tissue Lysate
Lane 2 : SK N BE (Human neuroblastoma) Whole Cell Lysate
Lane 3 : Human liver tissue lysate - total protein (ab29889)
Lane 4 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 2uM)
Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lane 6 : Liver (Rat) Tissue Lysate
Lane 7 : HeLa Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 8 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 9 : Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 10 : HeLa Whole Cell Lysate - Bleomycin Treated (20U/ml)
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 161 kDa
Observed band size: 171 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa, 95 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab66320 stained HeLa cells. The cells were 10% neutral buffered formalin fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66320, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab66320 staining ROCK2 in Human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66320, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
This product has been referenced in:
- Dalaklioglu S et al. Protective effect of exendin-4 treatment on erectile dysfunction induced by chronic methylglyoxal administration in rats. Peptides 106:1-8 (2018). IHC-P ; Rat . Read more (PubMed: 29792899) »
- Wang M et al. Oblongifolin M, an active compound isolated from a Chinese medical herb Garcinia oblongifolia, potently inhibits enterovirus 71 reproduction through downregulation of ERp57. Oncotarget 7:8797-808 (2016). Read more (PubMed: 26848777) »