ROCK1 Antibody - #AF7016

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製品: ROCK1 Antibody
カタログ: AF7016
タンパク質の説明: Rabbit polyclonal antibody to ROCK1
アプリケーション: WB IHC IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Sheep, Rabbit, Dog, Chicken
分子量: 160kDa; 158kD(Calculated).
ユニプロット: Q13464
RRID: AB_2835321

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説明書
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プロトコル
WB Handbook
IHC Protocol
IF/ICC Protocol

製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(91%)
クローナリティ:
Polyclonal
特異性:
ROCK1 Antibody detects endogenous levels of total ROCK1.
RRID:
AB_2835321
引用形式: Affinity Biosciences Cat# AF7016, RRID:AB_2835321.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

coiled-coil-containing protein kinase 1; coiled-coil-containing protein kinase I; MGC131603; MGC43611; p160 Rhoassociated coiled coil-forming protein kinase; p160 ROCK-1; p160 ROCK1; p160ROCK; PRO0435; Renal carcinoma antigen NY REN 35; Renal carcinoma antigen NY-REN-35; Rho associated coiled coil containing protein kinase 1; Rho associated protein kinase 1; Rho kinase; Rho-alpha kinase; Rho-associated; Rho-associated protein kinase 1; ROCK I; ROCK-I; ROCK1; ROCK1_HUMAN; Rok; rokalpha;

免疫原

免疫原:
Uniprot:

Q13464(ROCK1_HUMAN) >>Visit HPA database.

遺伝子(ID):
ROCK1(6093)
発現特異性:
Q13464 ROCK1_HUMAN:

Detected in blood platelets.

タンパク質の説明:
ROCK1 Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of DAPK3, GFAP, LIMK1, LIMK2, MYL9/MLC2, PFN1 and PPP1R12A. Phosphorylates FHOD1 and acts synergistically with it to promote SRC-dependent non-apoptotic plasma membrane blebbing.
タンパク質配列:
MSTGDSFETRFEKMDNLLRDPKSEVNSDCLLDGLDALVYDLDFPALRKNKNIDNFLSRYKDTINKIRDLRMKAEDYEVVKVIGRGAFGEVQLVRHKSTRKVYAMKLLSKFEMIKRSDSAFFWEERDIMAFANSPWVVQLFYAFQDDRYLYMVMEYMPGGDLVNLMSNYDVPEKWARFYTAEVVLALDAIHSMGFIHRDVKPDNMLLDKSGHLKLADFGTCMKMNKEGMVRCDTAVGTPDYISPEVLKSQGGDGYYGRECDWWSVGVFLYEMLVGDTPFYADSLVGTYSKIMNHKNSLTFPDDNDISKEAKNLICAFLTDREVRLGRNGVEEIKRHLFFKNDQWAWETLRDTVAPVVPDLSSDIDTSNFDDLEEDKGEEETFPIPKAFVGNQLPFVGFTYYSNRRYLSSANPNDNRTSSNADKSLQESLQKTIYKLEEQLHNEMQLKDEMEQKCRTSNIKLDKIMKELDEEGNQRRNLESTVSQIEKEKMLLQHRINEYQRKAEQENEKRRNVENEVSTLKDQLEDLKKVSQNSQLANEKLSQLQKQLEEANDLLRTESDTAVRLRKSHTEMSKSISQLESLNRELQERNRILENSKSQTDKDYYQLQAILEAERRDRGHDSEMIGDLQARITSLQEEVKHLKHNLEKVEGERKEAQDMLNHSEKEKNNLEIDLNYKLKSLQQRLEQEVNEHKVTKARLTDKHQSIEEAKSVAMCEMEKKLKEEREAREKAENRVVQIEKQCSMLDVDLKQSQQKLEHLTGNKERMEDEVKNLTLQLEQESNKRLLLQNELKTQAFEADNLKGLEKQMKQEINTLLEAKRLLEFELAQLTKQYRGNEGQMRELQDQLEAEQYFSTLYKTQVKELKEEIEEKNRENLKKIQELQNEKETLATQLDLAETKAESEQLARGLLEEQYFELTQESKKAASRNRQEITDKDHTVSRLEEANSMLTKDIEILRRENEELTEKMKKAEEEYKLEKEEEISNLKAAFEKNINTERTLKTQAVNKLAEIMNRKDFKIDRKKANTQDLRKKEKENRKLQLELNQEREKFNQMVVKHQKELNDMQAQLVEECAHRNELQMQLASKESDIEQLRAKLLDLSDSTSVASFPSADETDGNLPESRIEGWLSVPNRGNIKRYGWKKQYVVVSSKKILFYNDEQDKEQSNPSMVLDIDKLFHVRPVTQGDVYRAETEEIPKIFQILYANEGECRKDVEMEPVQQAEKTNFQNHKGHEFIPTLYHFPANCDACAKPLWHVFKPPPALECRRCHVKCHRDHLDKKEDLICPCKVSYDVTSARDMLLLACSQDEQKKWVTHLVKKIPKNPPSGFVRASPRTLSTRSTANQSFRKVVKNTSGKTS

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Sheep
100
Dog
100
Rabbit
100
Chicken
91
Zebrafish
73
Horse
0
Bovine
0
Xenopus
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13464 基板として

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
T3 Phosphorylation
K13 Ubiquitination
K65 Ubiquitination
K80 Ubiquitination
K109 Ubiquitination
K200 Ubiquitination
K225 Ubiquitination
Y255 Phosphorylation
S282 Phosphorylation
K294 Ubiquitination
K307 Ubiquitination
K339 Ubiquitination
T398 Phosphorylation
Y405 Phosphorylation
S407 Phosphorylation
K430 Ubiquitination
T455 Phosphorylation
S456 Phosphorylation
S533 Phosphorylation
K539 Ubiquitination
S567 Phosphorylation
T569 Phosphorylation
S572 Phosphorylation
S574 Phosphorylation
S576 Phosphorylation
Y603 Phosphorylation
Y604 Phosphorylation
K647 Acetylation
S679 Phosphorylation
S710 Phosphorylation
K718 Acetylation
S742 Phosphorylation
K782 Ubiquitination
K857 Ubiquitination
T887 Phosphorylation
T890 Phosphorylation
T897 Phosphorylation
K898 Ubiquitination
Y913 Phosphorylation
T917 Phosphorylation
K921 Ubiquitination
K950 Ubiquitination
T1024 Phosphorylation P41743 (PRKCI)
K1057 Ubiquitination
K1093 Ubiquitination
S1098 Phosphorylation
S1100 Phosphorylation
T1101 Phosphorylation
S1102 Phosphorylation
S1105 Phosphorylation
S1108 Phosphorylation
T1112 Phosphorylation
S1119 Phosphorylation
Y1153 Phosphorylation
T1180 Phosphorylation
T1189 Phosphorylation
K1194 Acetylation
Y1287 Phosphorylation
S1322 Phosphorylation
S1328 Phosphorylation
T1331 Phosphorylation
S1333 Phosphorylation P41743 (PRKCI)
T1334 Phosphorylation P41743 (PRKCI)
S1336 Phosphorylation
T1337 Phosphorylation P41743 (PRKCI)
S1341 Phosphorylation P41743 (PRKCI)

PTMs - Q13464 酵素として

Substrate Site Source
O00429 (DNM1L) S637 Uniprot
O14649 (KCNK3) S336 Uniprot
O14649 (KCNK3) S393 Uniprot
O14950 (MYL12B) T19 Uniprot
O14950 (MYL12B) S20 Uniprot
O14974 (PPP1R12A) T696 Uniprot
O14974 (PPP1R12A) S852 Uniprot
O14974 (PPP1R12A) T853 Uniprot
O43293 (DAPK3) T265 Uniprot
O43293 (DAPK3) T299 Uniprot
O60237 (PPP1R12B) T646 Uniprot
P04637 (TP53) S15 Uniprot
P07737 (PFN1) S138 Uniprot
P08670 (VIM) S72 Uniprot
P14136-1 (GFAP) T7 Uniprot
P14136 (GFAP) S13 Uniprot
P14136-1 (GFAP) S17 Uniprot
P14136 (GFAP) S38 Uniprot
P15311 (EZR) T567 Uniprot
P17661 (DES) S12 Uniprot
P17661 (DES) T17 Uniprot
P17661 (DES) S60 Uniprot
P17661 (DES) T76 Uniprot
P17661 (DES) T77 Uniprot
P19105 (MYL12A) T18 Uniprot
P19105 (MYL12A) S19 Uniprot
P19634 (SLC9A1) T653 Uniprot
P24844 (MYL9) T19 Uniprot
P24844 (MYL9) S20 Uniprot
P26038 (MSN) T558 Uniprot
P29966 (MARCKS) S159 Uniprot
P35241-1 (RDX) T564 Uniprot
P35241-1 (RDX) T573 Uniprot
P35611-1 (ADD1) T445 Uniprot
P35611-1 (ADD1) T480 Uniprot
P35611-3 (ADD1) T511 Uniprot
P48436 (SOX9) S181 Uniprot
P49815 (TSC2) T1203 Uniprot
P53667 (LIMK1) T508 Uniprot
P53671 (LIMK2) T505 Uniprot
P53671-1 (LIMK2) T526 Uniprot
P60484 (PTEN) S229 Uniprot
P60484 (PTEN) T232 Uniprot
P61587 (RND3) S7 Uniprot
P61587 (RND3) S11 Uniprot
P61587 (RND3) S218 Uniprot
P61587 (RND3) S240 Uniprot
Q05397 (PTK2) S732 Uniprot
Q14160 (SCRIB) S1378 Uniprot
Q14160 (SCRIB) S1508 Uniprot
Q16555-1 (DPYSL2) T555 Uniprot
Q8N264 (ARHGAP24) S391 Uniprot
Q8N264 (ARHGAP24) S402 Uniprot
Q8N264 (ARHGAP24) S413 Uniprot
Q8N264 (ARHGAP24) S415 Uniprot
Q8N264-1 (ARHGAP24) S437 Uniprot
Q8N264 (ARHGAP24) T452 Uniprot
Q8N264 (ARHGAP24) S573 Uniprot
Q8N264 (ARHGAP24) S574 Uniprot
Q8N264-1 (ARHGAP24) T575 Uniprot
Q8N264 (ARHGAP24) T576 Uniprot
Q8N264 (ARHGAP24) T577 Uniprot
Q9NRY4 (ARHGAP35) S1150 Uniprot
Q9NRY4 (ARHGAP35) T1173 Uniprot
Q9NRY4 (ARHGAP35) S1174 Uniprot
Q9NRY4 (ARHGAP35) T1226 Uniprot
Q9NRY4 (ARHGAP35) S1236 Uniprot
Q9UPT6 (MAPK8IP3) S314 Uniprot
Q9UPT6 (MAPK8IP3) S364 Uniprot
Q9UPT6 (MAPK8IP3) S365 Uniprot
Q9Y613 (FHOD1) S1131 Uniprot
Q9Y613 (FHOD1) S1137 Uniprot
Q9Y613 (FHOD1) T1141 Uniprot

研究背景

機能:

Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of DAPK3, GFAP, LIMK1, LIMK2, MYL9/MLC2, TPPP, PFN1 and PPP1R12A. Phosphorylates FHOD1 and acts synergistically with it to promote SRC-dependent non-apoptotic plasma membrane blebbing. Phosphorylates JIP3 and regulates the recruitment of JNK to JIP3 upon UVB-induced stress. Acts as a suppressor of inflammatory cell migration by regulating PTEN phosphorylation and stability (By similarity). Acts as a negative regulator of VEGF-induced angiogenic endothelial cell activation. Required for centrosome positioning and centrosome-dependent exit from mitosis (By similarity). Plays a role in terminal erythroid differentiation. May regulate closure of the eyelids and ventral body wall by inducing the assembly of actomyosin bundles (By similarity). Promotes keratinocyte terminal differentiation. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization (By similarity).

PTMs:

Autophosphorylated on serine and threonine residues.

Cleaved by caspase-3 during apoptosis. This leads to constitutive activation of the kinase and membrane blebbing.

細胞の位置付け:

Cytoplasm. Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome>Centriole. Golgi apparatus membrane>Peripheral membrane protein. Cell projection>Bleb. Cytoplasm>Cytoskeleton. Cell membrane. Cell projection>Lamellipodium. Cell projection>Ruffle.
Note: Associated with the mother centriole and an intercentriolar linker. Colocalizes with ITGB1BP1 and ITGB1 at the cell membrane predominantly in lamellipodia and membrane ruffles, but also in retraction fibers. Localizes at the cell membrane in an ITGB1BP1-dependent manner (By similarity). A small proportion is associated with Golgi membranes.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Detected in blood platelets.

サブユニット構造:

Homodimer. Interacts with RHOB, RHOC, MYLC2B and PTEN. Interacts with ITGB1BP1 (via N-terminus and PTB domain) (By similarity). Interacts with RHOA (activated by GTP), CHORDC1, DAPK3, GEM, JIP3, RHOE, PPP1R12A, PFN1, LIMK1, LIMK2 and TSG101. Interacts with FHOD1 in a Src-dependent manner. Interacts with SHROOM3.

タンパク質ファミリー:

The C-terminal auto-inhibitory domain interferes with kinase activity. RHOA binding leads to a conformation change and activation of the kinase. Truncated ROCK1 is constitutively activated.

Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.

研究領域

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Bacterial > Pathogenic Escherichia coli infection.

· Human Diseases > Infectious diseases: Bacterial > Shigellosis.

· Human Diseases > Infectious diseases: Bacterial > Salmonella infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Circulatory system > Vascular smooth muscle contraction.   (View pathway)

· Organismal Systems > Development > Axon guidance.   (View pathway)

· Organismal Systems > Immune system > Platelet activation.   (View pathway)

· Organismal Systems > Immune system > Leukocyte transendothelial migration.   (View pathway)

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

参考文献

1). β-patchoulene simultaneously ameliorated dextran sulfate sodium-induced colitis and secondary liver injury in mice via suppressing colonic leakage and flora imbalance. Biochemical Pharmacology, 2020 (PubMed: 33017576) [IF=5.8]

Application: WB    Species: mouse    Sample: colon

Fig.4.| Effect of β-PAE on TLR4/MyD88/NF-κB and ROCK/MLC signalling pathway.(A) Representative bands of TLR4, MyD88, phospho-IκBα, IκBα and p65. (B)Quantitative results of band densities of TLR4, MyD88, phospho-IκBα, IκBα and p65.(C) Representative bands of ROCK1, phospho-MLC2 and MLC2.

Application: WB    Species: mouse    Sample: Colons

Fig.4. |Effect of β-PAE on TLR4/MyD88/NF-κB and ROCK/MLC signalling pathway. (C) Representative bands of ROCK1, phospho-MLC2 and MLC2. (D) Quantitative results of band densities of ROCK1, phospho-MLC2 and MLC2.
2). Novel circRNA_0071196/miRNA‑19b‑3p/CIT axis is associated with proliferation and migration of bladder cancer. INTERNATIONAL JOURNAL OF ONCOLOGY, 2020 (PubMed: 32705161) [IF=5.2]

Application: WB    Species: Human    Sample: 5637 cells

Figure 11 Molecular basis of CIT action in BCa cells. Expression levels of p53, MLC2, CDK1, MDM2, CCND1 and ROCK1 mRNA (A) and protein (B and C) in sh-CIT and sh-Control-transfected 5637 cells. *P
3). Endostatin attenuates PDGF-BB- or TGF-β1-induced HSCs activation via suppressing RhoA/ROCK1 signal pathways. Drug Design Development and Therapy, 2019 (PubMed: 30666090) [IF=4.8]

Application: WB    Species: rat    Sample: HSC-T6 cells

Figure 4 |Endostatin inhibits the expression of α-SMA, RhoA, and ROCK1 at mRNA level.Notes: Transcript levels of α-SMA, RhoA, and ROCK1 were analyzed by RT-PCR (A–C). Endostatin significantly suppressed the expressions of α-SMA, RhoA, and ROCK1 at mRNA level in HSC-T6 cells. Data are expressed as mean ± SD. **P,0.01 (n=3 per group).
4). Experimental research on the evaluation of left ventricular systolic function by layered speckle tracking before and after berberine treatment in a cardiac hypertrophy rat model. Cardiovascular Diagnosis and Therapy, 2023 (PubMed: 37583682) [IF=2.4]

Application: IHC    Species: Rat    Sample:

Figure 5 Expression of ROCK protein in four groups of rats. Immunohistochemical staining, ×100. (A) Rats in group A; (B) rats in group B (positive, with a large amount of brown granular ROCK protein); (C) rats in group C; (D) rats in group D.
5). Brusatol-Enriched Brucea javanica Oil Ameliorated Dextran Sulfate Sodium-Induced Colitis in Mice: Involvement of NF-κB and RhoA/ROCK Signaling Pathways. Biomed Research International, 2021 (PubMed: 34414236)

Application: WB    Species: Mice    Sample: colon tissue

Figure 7 BE-BJO inhibited the activation of the RhoA/ROCK signaling pathway in DSS-induced UC in mice. Representative Western Blot images of GTP RhoA, total RhoA, ROCK-1, p-MLC, and MLC (a). The relative protein expressions of GTP RhoA (b), ROCK-1 (c), and p-MLC (d) in colon tissue were detected by Western Blot. All values are presented as the mean ± SEM. ##P < 0.01 versus normal group; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001 versus DSS group.
6). SKF38393 prevents high glucose (HG)-induced endothelial dysfunction by inhibiting the effects of HG on cystathionine γ-lyase/hydrogen sulfide activity and via a RhoA/ROCK1 pathway. Frontiers in Bioscience - Elite, 2022 (PubMed: 35226992)

Application: WB    Species: Human    Sample: HUVECs

Fig. 7. Activation of the DR1-CSE/H 2 S pathway attenuates HG-induced HUVEC dysfunction by inhibiting the RhoA/ROCK1 pathway. Analysis of p-RhoA, RhoA, p-ROCK1 and ROCK1 levels using western blot (n = 4). The intensity of each phosphorylated band was quantified by densitometry, and data was normalized to the corresponding total band signal. The results were expressed as the mean ± SEM. ** p < 0.01 vs. control group; # p < 0.05 vs. HG group; ## p < 0.01 vs. HG group; & p < 0.05 vs. HG+SKF38393 group; && p < 0.01 vs. HG+SKF38393 group.
7). Calycosin-7-O-β-D-Glucoside Treatment Promotes Axonal Regeneration via Rho/ROCK Pathway After Ischemia/Reperfusion Injury. Research Square, 2020

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