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H1152

Manufactured by Bio-Techne
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H1152 is a laboratory instrument designed for the analysis of various biological samples. It is a high-performance liquid chromatography (HPLC) system used for the separation, identification, and quantification of chemical compounds. The core function of H1152 is to provide reliable and accurate analytical data to support scientific research and product development.

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Lab products found in correlation

Blebbistatin, by Bio-Techne (4 mentions) Y 27632, by Bio-Techne (3 mentions) Y 27632, by Merck Group (3 mentions) Nocodazole, by Merck Group (3 mentions) Gm6001, by Merck Group (2 mentions) Blebbistatin, by Merck Group (2 mentions) Cytochalasin d, by Merck Group (2 mentions) Rock inhibitors y27632, by Bio-Techne (2 mentions) Integrin α6 cd49f antibody, by R&D Systems (2 mentions) Fak inhibitor 14, by Merck Group (2 mentions) Rs504393, by Bio-Techne (1 mentions) Axiovision microscope, by Zeiss (1 mentions) Transwell inserts with 8 μm pores, by BD (1 mentions) Tamoxifen citrate salt, by Merck Group (1 mentions) Fasudil, by Selleck Chemicals (1 mentions) Sulforhodamine b, by Merck Group (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Bca protein assay, by Thermo Fisher Scientific (1 mentions) Epigallocatechin gallate (egcg), by Bio-Techne (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Glycyl-H1152 dihydrochloride (3 citations)

16 protocols using h1152

1

Lung Endothelial Cell Transmigration Assay

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Primary lung endothelial cells (3x104) were seeded on gelatin-coated 24-well transwell inserts with 8μm pores (BD) and allowed to grow to confluency. Endothelial cells were pre-incubated with the following inhibitors: 50 μM RS504393 (CCR2), 50 μM 2-APB (IP3-gated Ca+ channels), 10 μM BAPTA-AM (chelating intracellular Ca2+), 1 μM ML-7 (MLCK), 5 μM H-1152 (ROCK2), 2.5 μM Blebbistatin (myosin II ATPase) for two hours in RPMI1640/10% FCS medium (all inhibitors were purchased from Tocris). The first four inhibitors were used according to manufacturer’s recommendations (Tocris); for H-1152 and Blebbistatin we used concentrations as previously published (28 (link)). Upon removal of inhibitors from the endothelial cells MC-38GFP cells (2x104) were added in presence or absence of CD115+ monocytes (1x105) in RPMI1640/3% FCS. Transmigration was induced with RPMI1640/10% FCS in the lower chamber and terminated after 16 hours. Alternatively, MC-38GFP cells (2x104) were added to the upper insert with or without rhCCL2 (1 μg/ml; kindly provided by A. Kungl, University of Graz). The number of transmigrated MC-38GFP cells was counted using a Zeiss AxioVision microscope (n ≥ 3).
Roblek M., Protsyuk D., Becker P.F., Stefanescu C., Gorzelanny C., Glaus Garzon J.F., Knopfova L., Heikenwalder M., Luckow B., Schneider S.W, & Borsig L. (2018). CCL2 is a vascular permeability factor inducing CCR2-dependent endothelial retraction during lung metastasis. Molecular cancer research : MCR, 17(3), 783-793.
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2

Kinase Inhibitor Screening Protocol

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AT13148 (Astex Pharmaceuticals), Y27632 (Tocris 1254), H1152 (Tocris 2414), fasudil (LC Laboratoris F-4660), GM6001 (Millipore 142880-36-2).
Rath N., Munro J., Cutiongco M.F., Jagiełło A., Gadegaard N., McGarry L., Unbekandt M., Michalopoulou E., Kamphorst J.J., Sumpton D., Mackay G., Vennin C., Pajic M., Timpson P, & Olson M.F. (2018). Rho kinase inhibitor AT13148 blocks pancreatic ductal adenocarinoma invasion and tumor growth. Cancer research, 78(12), 3321-3336.
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3

Cytoskeletal Modulation Agents Protocol

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Y27632 (Tocris 1254), H1152 (Tocris 2414), Fasudil (Selleckchem S1573, LC Laboratories F‐4660), Blebbistatin (Tocris 1760), GM6001 (Millipore 142880‐36‐2), Tamoxifen citrate salt (Sigma T9262), 4HT (4‐Hydroxytamoxifen, Sigma H7904).
Rath N., Morton J.P., Julian L., Helbig L., Kadir S., McGhee E.J., Anderson K.I., Kalna G., Mullin M., Pinho A.V., Rooman I., Samuel M.S, & Olson M.F. (2016). ROCK signaling promotes collagen remodeling to facilitate invasive pancreatic ductal adenocarcinoma tumor cell growth. EMBO Molecular Medicine, 9(2), 198-218.
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4

Analysis of MYPT1 and MLC20 Phosphorylation

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Reagents were obtained from the following sources: H1152, EGCG (Tocris Bioscience, Bristol, UK); protease inhibitor cocktail, sulforhodamine B, PMA, and 1α,25-dihydroxivitamin D3 (VD3) (Sigma-Aldrich, St. Louis, MO, USA); BCA protein assay (Thermo Fischer Scientific, Waltham, MA, USA). Antibodies and their sources were as follows: MYPT1pT696 and MYPT1pT853 (Millipore, MilliporeSigma, Burlington, MA, USA); horseradish-peroxidase conjugated anti-rabbit (Sigma-Aldrich, St. Louis, MO, USA); MYPT1 [45 (link)]; MLC20pS19, p21, PCNA, and horseradish-peroxidase conjugated anti-mouse (Cell Signaling Technology, Danvers, MA, USA); β-actin-HRP, PP1β, and CD11b (Santa Cruz Biotechnology, Dallas, TX, USA).
Tóth E., Erdődi F, & Kiss A. (2021). Myosin Phosphatase Is Implicated in the Control of THP-1 Monocyte to Macrophage Differentiation. International Journal of Molecular Sciences, 22(5), 2516.
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5

Apoptosis and Necrosis Induction Assay

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Cells were treated with 1 μM staurosporine for 4 hours (primary cells) or overnight (lymphoma cells) to induce apoptosis. Necrosis was induced by heat-killing at 60°C, for 30 minutes. Cell death was confirmed by flow cytometry detection of labeling with 7-Aminoactinomycin D (7-AAD) and Annexin V (Biolegend). Primary T cell activation was performed with 0.5 μg/ml anti-CD3 (OKT3, Thermo Fisher, 16-0037-85) + 0.5 μg/ml anti-CD28 (Thermo Fisher, 16-0288-85) in the presence of 500 IU IL-2 (Peprotech, UK). The following inhibitors were used at the concentrations indicated in the figures: blebbistatin (SigmaAldrich), H1152 (Tocris), Y27632 (Calbiochem), wortmannin (Calbiochem), latrunculin A (SigmaAldrich), cytochalasin D (Thermo Fisher Scientific).
Davies S.P., Reynolds G.M., Wilkinson A.L., Li X., Rose R., Leekha M., Liu Y.S., Gandhi R., Buckroyd E., Grove J., Barnes N.M., May R.C., Hubscher S.G., Adams D.H., Huang Y., Qureshi O, & Stamataki Z. (2019). Hepatocytes Delete Regulatory T Cells by Enclysis, a CD4+ T Cell Engulfment Process. Cell Reports, 29(6), 1610-1620.e4.
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6

Modulating Cytoskeletal Dynamics in Cells

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Blebs were generated using 14 μM Jas (Thermo Fisher, Invitrogen; Cat. No. J7473) for 15 min. Formin perturbation was carried out using 10–25 μM SMI-FH2 (Calbiochem; Cat. No. S4826-5MG) for 15 min–1 h based on experimental requirement. Arp2/3 inhibition was carried out using 200 μM CK-666 (Sigma-Aldrich; Cat. No. SML0006 5MG) treatment for 3 h. Ezrin perturbation was carried out using the inhibitor NSC668394 purchased from EMD Millipore (Cat. No. 341216-10MG). Cells were treated with 25 μM of the drug for 1 h. Myosin II perturbation was carried out using a cocktail of ML-7 (Sigma-Aldrich; Cat. No. I2764) and Y27632 (Sigma-Aldrich; Cat. No. Y0503-1MG) or H1152 purchased from Tocris (Cat. No. 2414). Cells were treated with a cocktail of the ML-7 and Y27632/ H1152 at a final concentration of 20 μM of each for 1 h. Owing to the reversible nature of the drugs acting on the target, imaging was carried out in the presence of the drug except in the case of Jas treatment. All drug treatments were carried out in HEPES buffer saline containing 2 mg/ml glucose at 37°C for the indicated time periods.
Sil P., Mateos N., Nath S., Buschow S., Manzo C., Suzuki K.G., Fujiwara T., Kusumi A., Garcia-Parajo M.F, & Mayor S. (2020). Dynamic actin-mediated nano-scale clustering of CD44 regulates its meso-scale organization at the plasma membrane. Molecular Biology of the Cell, 31(7), 561-579.
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7

Inhibition of Cytoskeletal Dynamics

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Nocodazole and cytochalasin D were purchased from Sigma Aldrich (St Louis, MO), and latrunculin A and swinholide A were gifts from Dr. Leah Haimo. Blebbistatin and ROCK inhibitors (Y27632 and H1152) were from Tocris Bioscience (Minnesota, USA). FAK inhibitor 14 was purchased from Sigma Aldrich (St Louis, MO) and Integrin α6/CD49f antibody was purchased from R&D systems (Minneapolis, MN).
Weng N.J., Cheung C, & Talbot P. (2018). Dynamic blebbing: A bottleneck to human embryonic stem cell culture that can be overcome by Laminin-Integrin signaling. Stem cell research, 33, 233-246.
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8

Inhibition of Cytoskeletal Dynamics

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Nocodazole and cytochalasin D were purchased from Sigma Aldrich (St Louis, MO), and latrunculin A and swinholide A were gifts from Dr. Leah Haimo. Blebbistatin and ROCK inhibitors (Y27632 and H1152) were from Tocris Bioscience (Minnesota, USA). FAK inhibitor 14 was purchased from Sigma Aldrich (St Louis, MO) and Integrin α6/CD49f antibody was purchased from R&D systems (Minneapolis, MN).
Weng N.J., Cheung C, & Talbot P. (2018). Dynamic blebbing: A bottleneck to human embryonic stem cell culture that can be overcome by Laminin-Integrin signaling. Stem cell research, 33, 233-246.
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9

Embryo Drug Injection and Actin Perturbation

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Embryos were prepared as described before. For drug injections, embryos were kept in box containing Drierite and injected with either drugs or similar volumes of their solvents (RNase free water or DMSO), at the end of cellularisation or early gastrulation. For ROCK inhibition, embryos were injected with 20mM H1152 (Tocris) and imaged immediately. For actin perturbations, embryos were injected with 5mM Lat A (Invitrogen) and imaged immediately. For anisotropy measurements, 15-20 Z sections separated by 0.25μm were acquired from the most apical membrane of the embryo.
Chandran R., Kale G., Philippe J.M., Lecuit T, & Mayor S. (2021). Distinct actin-dependent nanoscale assemblies underlie the dynamic and hierarchical organization of E-cadherin. Current biology : CB, 31(8), 1726-1736.e4.
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10

Quantifying HUVEC-HCT116 Cell Adhesion

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HUVEC cells were seeded in black wall clear bottom 96-well plates and allowed to adhere for 48 hours prior to each experiment. HUVEC cell monolayers were then starved in EBM incomplete media + 1% FBS for 1 hour and subsequently treated with TNFα (10 ng/ml) for 4 hours. HCT116 cells or HCT116 cells transfected with siRNA were starved in EBM incomplete + 1% FBS for 1 hour and then treated with CID16020046, CBD or with specific inhibitors targeting MEK1/2 (PD184161; Cayman) and ROCK (Y27632 [Sigma Aldrich, St. Louis, MO, USA] and H-1152 [Tocris]) for 1 hour at the indicated concentrations. Cells were washed once and loaded with calcein-AM (1:1000) in EBM incomplete + 1 % FBS for 15 min at 37°C in the dark. After two additional washing steps, HCT116 cells were added to the HUVEC monolayer and incubated for 30 min at 37°C at 90 rpm shaking. Cells were washed twice with HBSS before signals of adherent HCT116 cells were measured in a FLEX station II. Data are expressed as relative fluorescence units (RFU).
Kargl J., Andersen L., Hasenöhrl C., Feuersinger D., Stančić A., Fauland A., Magnes C., El‐Heliebi A., Lax S., Uranitsch S., Haybaeck J., Heinemann A, & Schicho R. (2015). GPR55 promotes migration and adhesion of colon cancer cells indicating a role in metastasis. British Journal of Pharmacology, 173(1), 142-154.
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