- PA5-22262 - Invitrogen Antibodies ROCK1 antibody

Zhang Y, Zhang X, Cao Z, Huang Y, Zheng Y, Yang X
Open life sciences 2021;16(1):1141-1150

Chen X, Tan QQ, Tan XR, Li SJ, Zhang XX
Cell death & disease 2021 Aug 26;12(9):809

Bolado-Carrancio A, Rukhlenko OS, Nikonova E, Tsyganov MA, Wheeler A, Garcia-Munoz A, Kolch W, von Kriegsheim A, Kholodenko BN
eLife 2020 Jul 24;9

Thompson JM, Nguyen QH, Singh M, Pavesic MW, Nesterenko I, Nelson LJ, Liao AC, Razorenova OV
Oncogene 2017 Feb 23;36(8):1080-1089

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of ROCK1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: ROCK1 Polyclonal Antibody (Product # PA5-22262) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ROCK1 antibody immunoprecipitates ROCK1 protein in IP experiments. IP Sample: A431 whole cell lysate/extract A : Control with 3 µg of pre-immune rabbit IgG B : Immunoprecipitation of ROCK1 by 3 µg of ROCK1 antibody (Product # PA5-22262) 5% SDS-PAGE The immunoprecipitated ROCK1 protein was detected by ROCK1 antibody (Product # PA5-22262) diluted at 1 : 500. Anti-rabbit IgG (HRP) was used as a secondary reagent.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Synthetic lethality of Y-27632 with VHL loss is mimicked by siRNA downregulation of ROCK1, not ROCK2 RCC4+-VHL matched cell lines were transfected with siRNAs targeting ROCK1, ROCK2, or non-targeting siRNA control (siControl). Twenty-four hours after transfection cells were plated for a clonogenic assay. Each transfection was done in triplicate, followed by clonogenic assays conducted in triplicate, and the experiments were repeated at least two times. ( a ) Transfection with siROCK1, but not siROCK2, resulted in significant reduction in RCC4 colony numbers in comparison to RCC4VHL. Thus, ROCK1 downregulation mimics the effect of Y-27632 treatment on viability of RCC4 cells, making it a likely target for Y-27632 causing synthetic lethality effect. Statistical analysis was performed using a paired t-test comparing numbers of colonies in each siROCK group to siControl. SEMs are shown. (b-c) The degree of each target knockdown by its specific siRNA (as indicated) was assessed by Western blot.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 2 miR-206 inhibited lipogenesis and TG secretion in vivo. A Representative images of H&E staining (upper), oil-red o staining (middle), and IHC staining (bottom) in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. B Secreted TG levels in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. C miR-206 levels in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. D ROCK1 mRNA levels in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. E , F ROCK1/AMPK activities in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. G Protein levels of ROCK1, p-AMPK, and lipogenesis-related proteins in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. H Quantifications of G . * p < 0.05, ** p < 0.01, and *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Overexpression of AOC3 increases the ROCK1 expression and phosphorylation of MLC 20 in RIMSCs under hypoxia. ROCK1 and p-MLC 20 protein expression in RIMSCs were determined by western blotting. GAPDH and MLC 20 were used as internal controls, respectively (* P < 0.05, ** P < 0.01, and *** P < 0.001).

PA5-22262