ROCK1 Antibody (154C1465) - (active/cleaved) - BSA Free

Western Blot: cleaved ROCK1 Antibody (154C1465) [NB100-56596] - Analysis using Azide Free version of NB100-56596. Cleaved ROCK-1 in A) untreated, B) 1 hour and C) 4 hour staurosporin-treated (2 uM) Jurkat cells using this antibody. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used

• Reactivity: Hu, Mu, Rt, Ca, Pm, Rb
• Applications: WB, ICC/IF
• Clone: 154C1465
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

ROCK1 Antibody (154C1465) - (active/cleaved) - BSA Free Summary

• Immunogen: This antibody was generated by immunizing mice with a synthetic peptide sequence corresponding to amino acids 1114-1126 GNLPESRIEGWLS of human ROCK-1. This sequence is 100% conserved between human, chimpanzee, mouse, rat, dog, and rabbit. This immunogen peptide sequence is adjacent to the caspase-3 cleavage site of ROCK-1.
• Specificity: This mAb recognizes the cleaved C-terminus of ROCK-1. It does not recognize the full-length protein. Since ROCK-1 is cleaved by caspase-3 during apoptosis, this may be used for detection of apoptosis.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Mouse
• Gene: ROCK1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence reported in scientific literature (PMID 21059388)
  • Western Blot 1-2 ug/ml

Reactivity Notes

Human reactivity reported in scientific literature (PMID:32861242). Cross reacts with Chimpanzee.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for ROCK1 Antibody (154C1465) - (active/cleaved) - BSA Free

• EC 2.7.11
• EC 2.7.11.1
• MGC131603
• MGC43611
• p160 ROCK-1
• p160ROCK
• p160-ROCK
• PRO0435
• Renal carcinoma antigen NY-REN-35
• Rho kinase
• rho-associated protein kinase 1
• Rho-associated, coiled-coil containing protein kinase 1
• Rho-associated, coiled-coil-containing protein kinase 1
• ROCK1
• ROK beta

Background

The Rho target protein, Rho-associated coiled coil-containing protein kinase (ROCK)-1 has been found to be a new Caspase-3 substrate. ROCK, one of the effectors of the small GTPase Rho, has recently been shown to contribute significantly to myosin light chain (MLC) activation through two pathways: direct phosphorylation of MLC and phosphorylation of MLC phosphatase, leading to its inhibition. The increase in cellular contractility that is necessary for apoptotic membrane blebbing implies sustained augmentation of MLC phosphorylation. ROCK-1 consists of an amino-terminal kinase domain and an inhibitory cysteine/histidine-rich C-terminal domain that is located within a pleckstrin-homology region. They are joined by a variable region that contains the Rho-binding domain. During apoptosis, ROCK-I is cleaved by caspase-3 at a conserved DETD1113/G sequence and its carboxy-terminal inhibitory domain is removed, resulting in deregulated and constitutive kinase activity. The caspase-3-mediated cleavage and activation of ROCK I induces phosphorylation of MLC and membrane blebbing, one of the first events in the execution phase of apoptosis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for cleaved ROCK1 Antibody (NB100-56596)(5)

Publications using NB100-56596

• McKenzie BA, Fernandes JP, Doan MAL et al. Activation of the executioner caspases-3 and -7 promotes microglial pyroptosis in models of multiple sclerosis J Neuroinflammation 2020-08-29 [PMID: 32861242] (ICC/IF, WB, Human)
• Goetsch K P, Kallmeyer K, Niesler C U. Decorin modulates collagen I-stimulated, but not fibronectin-stimulated, migration of C2C12 myoblasts. Matrix Biol. 2011-03-01 [PMID: 21059388] (ICC/IF)
• Orlando KA, Pittman RN. Rho kinase regulates phagocytosis, surface expression of GlcNAc, and Golgi fragmentation of apoptotic PC12 cells. Exp Cell Res. 2006-10-15 [PMID: 16904666]
  
  Details:
  Products cited: 1. ROCK-1 (Cleaved), IMG-383A: WB, Figs 1,2 (serum-starved PC12 cells). IMG-383 detected a 30 kDa cleaved Rock-1 band in serum-starved PC12 cells. The addition of the Broad-Spectrum Caspase Inhibitor Z-VAD-FMK (IMI-2310-1, -5) during serum
• Orlando KA, Stone NL, Pittman RN. Rho kinase regulates fragmentation and phagocytosis of apoptotic cells. Exp Cell Res. 2006-01-01 [PMID: 16259978] (WB)
  
  Details:
  Products cited: 1. ROCK-1 (Cleaved), IMG-383A: WB, Fig 3 (tamoxifen-treated Cos7 cells). IMG-383 detected a 30 kDa cleaved Rock-1 band in tamoxifen-treated cells. The addition of the Broad-Spectrum Caspase Inhibitor Z-VAD-FMK (IMI-2310-1, -5) prior to tam
• Zhao H, Miller M, Pfeiffer K et al. Anoxia and reoxygenation of human endothelial cells decrease ceramide glucosyltransferase expression and activates caspases. FASEB J. 2003-04-01 [PMID: 12586734] (WB)
  
  Details:
  WB (Normoxic/anoxic/reoxygenated HUVEC cells), Fig 1.

Review for cleaved ROCK1 Antibody (NB100-56596) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 08/01/2018

Summary

• Application: Western Blot
• Sample Tested: Adult brain
• Species: Human
• Lot: 121801A03

Bioinformatics

• Gene Symbol: ROCK1
• Entrez:
  • 6093(Human)
  • 19877(Mouse)
  • 81762(Rat)
• Uniprot:
  • O60259()