- PA5-78290 - Invitrogen Antibodies ROCK2 antibody

Submitted references Lamin A and the LINC complex act as potential tumor suppressors in Ewing Sarcoma.

Chiarini F, Paganelli F, Balestra T, Capanni C, Fazio A, Manara MC, Landuzzi L, Petrini S, Evangelisti C, Lollini PL, Martelli AM, Lattanzi G, Scotlandi K
Cell death & disease 2022 Apr 14;13(4):346

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ROCK2 in A) Neuro2A whole cell lysate, B) GL261 whole cell lysate, C) C8D30 whole cell lysate, D) NIH-3T3 whole cell lysate, E) BCL-1 whole cell lysate, F) Raw 264.7 whole cell lysate, G) C2Cl2 whole cell lysate using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with ROCK2 polyclonal antibody (Product # PA5-78290) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ROCK2 in A) A549 whole cell lysate, B) H1299 whole cell lysate, C) HCT116 whole cell lysate using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with ROCK2 polyclonal antibody (Product # PA5-78290) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ROCK2 in non-transfected (-) and transfected (+) 293T cells using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with ROCK2 polyclonal antibody (Product # PA5-78290) using a dilution of 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ROCK2 in whole cell lysate using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with ROCK2 polyclonal antibody (Product # PA5-78290) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ROCK2 in Rat2 whole cell lysate using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with ROCK2 polyclonal antibody (Product # PA5-78290) using a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with ROCK2 Polyclonal Antibody (Product # PA5-78290) diluted at a dilution of 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein by western blot analysis. A. 30 µg Neuro2A whole cell lysate/extract. B. 30 µg GL261 whole cell lysate/extract. C. 30 µg C8D30 whole cell lysate/extract. D. 30 µg NIH-3T3 whole cell lysate/extract. E. 30 µg BCL-1 whole cell lysate/extract. F. 30 µg Raw 264.7 whole cell lysate/extract. G. 30 µg C2Cl2 whole cell lysate/extract.5 % SDS-PAGE. ROCK2 Polyclonal Antibody (Product # PA5-78290) dilution: 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with ROCK2 Polyclonal Antibody (Product # PA5-78290) diluted at a dilution of 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein by western blot analysis. A. 30 µg A549 whole cell lysate/extract. B. 30 µg H1299 whole cell lysate/extract. C. 30 µg HCT116 whole cell lysate/extract.5 % SDS-PAGE. ROCK2 Polyclonal Antibody (Product # PA5-78290) dilution: 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein by western blot analysis. A. 30 µg Neuro2A whole cell lysate/extract. B. 30 µg GL261 whole cell lysate/extract. C. 30 µg C8D30 whole cell lysate/extract. D. 30 µg NIH-3T3 whole cell lysate/extract. E. 30 µg BCL-1 whole cell lysate/extract. F. 30 µg Raw 264.7 whole cell lysate/extract. G. 30 µg C2Cl2 whole cell lysate/extract.5 % SDS-PAGE. ROCK2 Polyclonal Antibody (Product # PA5-78290) dilution: 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein by western blot analysis. A. 30 µg Rat2 whole cell lysate/extract.5 % SDS-PAGE. ROCK2 Polyclonal Antibody (Product # PA5-78290) dilution: 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of ROCK2 was achieved by transfecting HeLa with ROCK2 specific siRNAs (Silencer® select Product # s18162). Western blot analysis (Fig. a) was performed using whole cell extracts from the ROCK2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ROCK2 Polyclonal Antibody (Product # PA5-78290, 1:3000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to ROCK2. An uncharacterized band around 53 kDa was also observed.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-ROCK2 Polyclonal Antibody (Product # PA5-78290) and a 180 kDa corresponding to ROCK2 along with an uncharacterized band around 53 kDa was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), MCF-7 (Lane 3), MDA-MB-231 (Lane 4), U-87 MG (Lane 5) and C1C12 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by XCell SureLock™ Mini-Cell and XCell II™ Blot Module (Product # EI0002). The blot was probed with the primary antibody (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein at cytoplasm by immunofluorescent analysis. Sample: A431 cells were fixed in ice-cold MeOH for 5 min. Green: ROCK2 protein stained by ROCK2 Polyclonal Antibody (Product # PA5-78290) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein at cytoplasm by immunofluorescent analysis. Sample: A431 cells were fixed in ice-cold MeOH for 5 min. Green: ROCK2 protein stained by ROCK2 Polyclonal Antibody (Product # PA5-78290) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ROCK2 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ROCK2 Rabbit Polyclonal Antibody (Product # PA5-78290) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein expression by immunohistochemical analysis. Sample: Frozen-sectioned adult mouse cerebellum. Green: ROCK2 protein stained by ROCK2 Polyclonal Antibody (Product # PA5-78290) diluted at 1:250. Red: NF-H, stained by NF-H antibody [GT114] diluted at 1:500. Blue: Fluoroshield with DAPI . Antigen Retrieval: Citrate buffer, pH 6.0, 10 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein expression by immunohistochemical analysis. Sample: Frozen-sectioned adult mouse cerebellum. Green: ROCK2 protein stained by ROCK2 Polyclonal Antibody (Product # PA5-78290) diluted at 1:250. Red: NF-H, stained by NF-H antibody [GT114] diluted at 1:500. Blue: Fluoroshield with DAPI . Antigen Retrieval: Citrate buffer, pH 6.0, 10 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ROCK2 Polyclonal Antibody detects ROCK2 protein at cytosol on mouse kidney by immunohistochemical analysis. Sample: Paraffin-embedded mouse kidney. ROCK2 Polyclonal Antibody (Product # PA5-78290) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: The expression of lamin A affects YAP and ROCK2 activity and stimulates neural differentiation in EWS cells. a Lamin A-GFP (green) and YAP (red) localization in Empty-GFP clone (pEV #2) and lamin A-GFP #84 (#84). DNA was counterstained with DAPI (DAPI). Merge of fluorescence signals are shown (MERGE). Graphs indicate the fluorescence intensity profile along the white arrows. Representative graphs of at least 30 nuclei analyzed for each sample were shown; magnification 100x, scale bar 10 mum; b Western blotting analysis of YAP, p(Ser127) YAP and MYC protein expression in Empty-GFP clone (pEV #2) and lamin A-GFP #84 (#84). GAPDH was used as loading control. Densitometric analysis is shown as mean values +- SD of three different experiments. Asterisks indicate statistically significant differences with respect to Empty-GFP clone; two-tailed unpaired Student's t-test, * p < 0.05; c Western blotting analyses of lamin A/C and ROCK2 protein expression in Empty-GFP clone (pEV #2), lamin A-GFP #30-40 (#30-40) and lamin A-GFP #84 (#84). GAPDH was used as loading control. Densitometric analysis is shown as mean values +- SD of three different experiments. Asterisks indicate statistically significant differences with respect to Empty-GFP clone; one-way ANOVA test, * p < 0.05, *** p < 0.001; d Western blotting analysis of lamin A/C and ROCK2 protein expression in siRNA scramble cells (SCR) and siLMNA A-673 ( siLMNA ). GAPDH was used as loading control. Densitometric analysis is shown as m

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Mevinolin induces neural differentiation and rescues YAP and ROCK2 dynamics in EWS cells. a Lamin A/C (green) and beta3-tubulin (red) localization in non-treated TC-71 cells (CTRL) and in mevinolin treated (5 muM) EWS cells (MEV). DNA was counterstained with DAPI (DAPI). Merge of fluorescence signals are shown (MERGE); magnification 100x, scale bar 10 mum; b Lamin A/C (green) and neurofilament-H (NEF-H) (red) localization in non-treated TC-71 cells (CTRL) and in mevinolin treated (5 muM) EWS cells (MEV). DNA was counterstained with DAPI (DAPI). Merge of fluorescence signals are shown (MERGE); magnification 100x, scale bar 10 mum; c qRT-PCR analysis of LMNA , NEF-H , beta3-tubulin, nestin and SOX2 genes in untreated TC-71 cells (CTRL) and in mevinolin treated EWS cells (2.5 muM or 5muM MEV). Data are shown as 2- DeltaDelta Ct . GAPDH was used as a housekeeping gene. Data are shown as mean values +- SD of three different experiments. Asterisks indicate statistically significant differences with respect to CTRL cells; one-way ANOVA test, * p < 0.05, ** p < 0.01, *** p < 0.001; d Lamin A/C (green) and YAP (red) localization in non-treated TC-71 cells (CTRL) and in mevinolin treated (5 muM) EWS cells (MEV). DNA was counterstained with DAPI (DAPI). Merge of fluorescence signals are shown (MERGE). Graphs indicate the fluorescence intensity profile along the white arrows. Representative graphs of at least 30 nuclei analyzed for each sample were shown; magnification 100x, scale bar 10

PA5-78290

Antibody data