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ROCK1 Recombinant antibody

ROCK1 Uni-rAbTM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

230423C6

Cat no : 83166-5-RR

Synonyms

Renal carcinoma antigen NY-REN-35, P160ROCK, p160 ROCK-1, p160 ROCK 1, EC:2.7.11.1

Validation Data Gallery

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  • Western Blot (WB) analysis of various lysates using ROCK1 Recombinant antibody (83166-5-RR)

    WB analysis using 83166-5-RR

    Various lysates were subjected to SDS PAGE followed by western blot with 83166-5-RR (ROCK1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of various lysates using ROCK1 Recombinant antibody (83166-5-RR)

    WB analysis using 83166-5-RR

    Various lysates were subjected to SDS PAGE followed by western blot with 83166-5-RR (ROCK1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

  • Immunohistochemistry (IHC) staining of human intrahepatic cholangiocarcinoma tissue using ROCK1 Recombinant antibody (83166-5-RR)

    IHC staining of human intrahepatic cholangiocarcinoma using 83166-5-RR

    Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 83166-5-RR (ROCK1 antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunofluorescence (IF) / fluorescent staining of HeLa cells using ROCK1 Recombinant antibody (83166-5-RR)

    IF Staining of HeLa using 83166-5-RR

    Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using ROCK1 antibody (83166-5-RR, Clone: 230423C6 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1).

  • Flow cytometry (FC) experiment of HeLa cells using ROCK1 Recombinant antibody (83166-5-RR)

    FC experiment of HeLa using 83166-5-RR

    1x10^6 HeLa cells were intracellularly stained with 0.25 ug ROCK1 Recombinant antibody (83166-5-RR, Clone:230423C6) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

  • Affinity and Kinetic Characterization of 83166-5-RR

    Affinity and Kinetic Characterization of 83166-5-RR

    Biolayer interferometry (BLl) kinetic assays of 83166-5-RR against Human ROCK1 were performed. The affinity constant is 0.401 nM.

Tested Applications

Positive WB detected inJurkat cells, NIH/3T3 cells, HeLa cells, A431 cells, RAW 264.7 cells
Positive IHC detected inhuman intrahepatic cholangiocarcinoma tissue
Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
Positive IF/ICC detected inHeLa cells
Positive FC (Intra) detected inHeLa cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunohistochemistry (IHC)IHC : 1:50-1:500
Immunofluorescence (IF)/ICCIF/ICC : 1:125-1:500
Flow Cytometry (FC) (INTRA)FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 83166-5-RR is ROCK1 Fusion Protein expressed in E. coli.

Tested Reactivity human, mouse
Host / Isotype Rabbit / IgG
Class Recombinant
Type Antibody
Immunogen ROCK1 fusion protein Ag16317
Full Name Rho-associated, coiled-coil containing protein kinase 1
Calculated Molecular Weight 1354 aa, 158 kDa
Observed Molecular Weight 150-160 kDa
GenBank Accession NumberBC113114
Gene Symbol ROCK1
Gene ID (NCBI) 6093
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purfication
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.

Background Information

ROCK1, also named as p160ROCK and NY-REN-35, belongs to the protein kinase superfamily and AGC Ser/Thr protein kinase family. ROCK1 phosphorylates and activates DAPK3, which then regulates myosin light chain phosphatase through phosphorylation of MYPT1 thereby regulating the assembly of the actin cytoskeleton, cell migration, invasiveness of tumor cells, smooth muscle contraction and neurite outgrowth. ROCK1 is required for centromere positioning and centromere-dependent exit from mitosis. It is necessary for apoptotic membrane blebbing. ROCK1 catalyzes the reaction: ATP + a protein = ADP + a phosphoprotein.

Protocols

Product Specific Protocols
WB protocol for ROCK1 antibody 83166-5-RRDownload protocol
IHC protocol for ROCK1 antibody 83166-5-RRDownload protocol
IF protocol for ROCK1 antibody 83166-5-RRDownload protocol
FC protocol for ROCK1 antibody 83166-5-RRDownload protocol
Standard Protocols
Click here to view our Standard Protocols