ROCK1 Recombinant antibody

ROCK1 Uni-rAb^TM 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

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Validation Data Gallery

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  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.

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  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.

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  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).

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  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).

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  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).

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  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.

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Tested Applications

• Positive WB detected in: Jurkat cells, NIH/3T3 cells, HeLa cells, A431 cells, RAW 264.7 cells
• Positive IHC detected in: human 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 in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/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 Number: BC113114
• Gene Symbol: ROCK1
• Gene ID (NCBI): 6093
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC 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.