Y 27632

Manufactured by Apexbio

Sourced in United States

Y-27632 is a small molecule inhibitor that selectively and potently inhibits the Rho-associated protein kinase (ROCK) enzymes ROCK1 and ROCK2. It is commonly used as a research tool to study the role of ROCK signaling in various biological processes.

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Lab products found in correlation

Wistar kyoto wky rats, by Charles River Laboratories (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Ccg 100602, by Apexbio (2 mentions) Alzet osmotic minipumps model 2ml2, by Durect (2 mentions) A83 01, by Merck Group (2 mentions) Collagenase type 4, by Worthington (2 mentions) Transwell insert, by Corning (2 mentions) Nicotinamide, by Merck Group (2 mentions) Murine egf, by Thermo Fisher Scientific (2 mentions) Primocin, by InvivoGen (2 mentions) N acetyl cysteine (nac), by MP Biomedicals (2 mentions) Rat tail collagen type 1, by Corning (2 mentions) 12 well polystyrene tissue culture dishes, by Corning (2 mentions) N n 3 5 difluorophenacetyl l alanyl s phenylglycine t butyl ester dapt, by Apexbio (2 mentions) Human collagen type 1, by Advanced BioMatrix (2 mentions) Gastrin, by AnaSpec (2 mentions) Sb202190, by Selleck Chemicals (2 mentions) Matrigel, by Corning (2 mentions) Fluorescein diacetate, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions)

Spelling variants of this product

ROCK inhibitor Y-27632 (4 citations) Y-27632 dihydrochloride (2 citations)

19 protocols using y 27632

OVA-induced Airway Hyperreactivity Modulation

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Animals were randomly divided into three groups (n = 10 for each group): OVA group, animals received OVA aerosol using sensitization protocol; OVA+Y-27632 group, animals were subjected to inhalation of Rho-kinase inhibitor (1mM) (Y-27632, Apexbio, USA) 10 min before the eight last OVA exposures for 6 min; control group, animals only received saline aerosol as the same protocol used for OVA administration. In addition, there was an OVA+saline group in which animals received saline before OVA administrations similar to the protocol used for the application of Rho-kinase inhibitor in OVA+Y-27632 group.
Since changes in airway conductance and eosinophil infiltration reaches to the maximum in 72h following antigen exposure in Guiana pigs, we examined AHR and inflammation 72h after the last antigen challenge [31 (link)–33 (link)].

Pazhoohan S., Raoufy M.R., Javan M, & Hajizadeh S. (2017). Effect of Rho-kinase inhibition on complexity of breathing pattern in a guinea pig model of asthma. PLoS ONE, 12(10), e0187249.

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Spontaneously Hypertensive Rat Drug Treatments

Cited 1 time

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Adult (4 month-old) male spontaneously hypertensive rats (SHR) and normotensive control Wistar-Kyoto (WKY) rats (Charles River Laboratories, San Diego, CA) were studied as described previously [10 (link)]. All animal procedures were performed in accordance with NIH guidelines (Guide for the Care and Use of Laboratory Animals, revised 2011) and protocols approved by the Institutional Animal Care and Use Committee of Loma Linda University. For in vivo drug treatments, Y-27632 (0.3 mg/kg/h, ApexBio Technology, TX), CCG-100602 (7.5 mg/kg/d, ApexBio Technology, TX) or vehicle control (DMSO, Sigma-Aldrich) were continuously administered for 2 weeks by Alzet osmotic minipumps (Model 2ML2, DURECT, CA), implanted subcutaneously in rats under anesthesia with 2% isoflurane (JD Medical, AZ) [10 (link)].

Zhou N., Lee J.J., Stoll S., Ma B., Costa K.D, & Qiu H. (2017). Rho Kinase Regulates Aortic Vascular Smooth Muscle Cell Stiffness Via Actin/SRF/Myocardin in Hypertension. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 44(2), 701-715.

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Hepatocyte Culture Optimization Protocol

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Rat tail type I collagen, Transwell inserts (0.9 cm² cell culture area, 1.6 × 10⁶ pores/cm²), 12-well polystyrene tissue culture dishes, 12-well polycarbonate plates and Transwell inserts (1.12 cm² cell culture area, 1.6 × 10⁸ pores/cm²) were purchased from Corning (Corning, NY). SB202190 was from Selleckchem (Houston, TX). Collagenase (type IV) was purchased from Worthington Biochemicals (Lakewood, NJ). Y-27632 and N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) were purchased from ApexBio Technology (Houston, TX). Human collagen (type I) was obtained from Advanced Biomatrix (Carlsbad, CA). The ALP assay kit was from AbCam (Cambridge, MA). Gastrin was from Anaspec (Freemont, CA). UGT-Glo™ Assay and P450-Glo™ CYP3A4 Assay were obtained from Promega (Madison, WI). N-acetyl cysteine was purchased from MP Biomedicals (Santa Ana, CA). Murine EGF was procured from Peprotech (Rock Hill, NJ). Primocin was purchased from InvivoGen (San Diego, CA). Nicotinamide, L-alanine p-nitroanilide, fluorescein diacetate, loperamide, zafirlukast, cobalt chloride, potassium phosphate buffer, bovine serum albumin, and A83–01 were acquired from Sigma-Aldrich (St. Louis, MO). All other reagents and Bicinchoninic Acid (BCA) Protein Assay Kit were purchased from Thermo Fisher Scientific (Waltham, MA).

Speer J.E., Wang Y., Fallon J.K., Smith P.C, & Allbritton N.L. (2019). Evaluation of human primary intestinal monolayers for drug metabolizing capabilities. Journal of Biological Engineering, 13, 82.

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RSV infection of A549 and primary nasal epithelial cells

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Recombinant wild-type RSV strain A2 in which enhanced GFP was inserted between the P and M genes was propagated, sucrose-purified, and titered by plaque assay on Vero cells, as previously described (Munir et al., 2008 (link)). The virus stock was sequenced and had no adventitious mutations, as confirmed by Sanger dideoxy sequencing. A549 cells were seeded and transfected with siRNA to MDA5, RIG-I, MAVS, or a nonsilencing negative control as described for HRV infections. Alternatively, primary nasal epithelial cells were digested from feeder cells and seeded in 12-well plates at 150,000 cells per well in 1 ml epithelial culture medium (Promocell) with 10 µM Y-27632 (ApexBio) and incubated at 37°C in 5% CO2. The cells were seeded on plates previously coated with 300 µl rat tail collagen (BD) at 30 µg/ml in PBS for 45 min at room temperature, washed twice with PBS, and air dried for 20 min. 36 h later, the cells were washed once with PBS. Transfected A549 cells or primary nasal epithelial cells were infected with RSV-GFP at an MOI of 1 or 0.2, respectively (diluted in appropriate complete medium, 300 µl per 12-well), for 1 h at room temperature, and washed twice with PBS to remove unadsorbed virus, and then medium was replaced (complete F-12K for A549; epithelial culture media without Y-27632 for nasal epithelial cells) and cultures were returned to 37°C in 5% humidified CO_2.

Lamborn I.T., Jing H., Zhang Y., Drutman S.B., Abbott J.K., Munir S., Bade S., Murdock H.M., Santos C.P., Brock L.G., Masutani E., Fordjour E.Y., McElwee J.J., Hughes J.D., Nichols D.P., Belkadi A., Oler A.J., Happel C.S., Matthews H.F., Abel L., Collins P.L., Subbarao K., Gelfand E.W., Ciancanelli M.J., Casanova J.L, & Su H.C. (2017). Recurrent rhinovirus infections in a child with inherited MDA5 deficiency. The Journal of Experimental Medicine, 214(7), 1949-1972.

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Scalp Keratinocyte Isolation and Culture

Cited 1 time

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In accordance with the Declaration of Helsinki Principles, ethical approval was
granted by the Institute of Ethical Committee of Nanfang Hospital, Southern
Medical School (approval number: NFEC-2021-265). HPKs were derived from the
scalp tissues of 20 to 30-year-old men or women after dermatoplasty. Personal
identity information was not obtained. To facilitate enzymatic lysis, the skin
was cut into pieces and incubated overnight in dispase II (Roche, Mannheim,
Germany) at 4°C. The next day, the epidermis was digested with 0.25%
trypsin-EDTA solution (Coolaber, China) for 20 min at 37°C, as previously
described.^{19 (link)} The isolated keratinocytes were cultured in defined
keratinocyte-SFM (ScienCell, 2111, USA) plus 20 μM Y-27632 (APExBIO, B1293,
Houston, USA),²⁰ and the medium was changed every 3 days.

He S., Wu H., Huang J., Li Q., Huang Z., Wen H, & Li Z. (2023). 3-D tissue-engineered epidermis against human primary keratinocytes apoptosis via relieving mitochondrial oxidative stress in wound healing. Journal of Tissue Engineering, 14, 20417314231163168.

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hPSC Single-Cell Dissociation and FACS

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1 ml Accutase (Sigma, A6964) was added to each well of hPSC cultures in a 6-well plate and incubated at 37°C for 5-10 min, until cells detached. An equal volume of mTeSR Plus medium (StemCell Technologies, 100-0276) with 10 µM Y-27632 (APExBIO, B1293) was added to cells and cells were dissociated mechanically by pipetting with a P1000 pipette twice then centrifuged at 300 g for 5 min at 4°C. Excess medium was removed and cells were resuspended in fluorescence-activated cell sorting (FACS) buffer (1× PBS containing 2% BSA, 10 µM Y-27632 and 100 U/ml penicillin-streptomycin). Cells were passed through a 70 µm cell strainer, pre-coated with FACS buffer, and centrifuged at 300 g for 5 min at 4°C. Cells were resuspended in 1 ml FACS buffer and transferred to 5 ml FACS tubes (Corning, 352063). DAPI (0.2 µg/ml) was added to respective tubes. Flow cytometry was performed using a Bio Rad Ze5#3 and accompanying software.

Hein R.F., Conchola A.S., Fine A.S., Xiao Z., Frum T., Brastrom L.K., Akinwale M.A., Childs C.J., Tsai Y.H., Holloway E.M., Huang S., Mahoney J., Heemskerk I, & Spence J.R. (2022). Stable iPSC-derived NKX2-1+ lung bud tip progenitor organoids give rise to airway and alveolar cell types. Development (Cambridge, England), 149(20), dev200693.

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Spontaneously Hypertensive Rat Drug Treatments

Cited 1 time

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Characterization of Breast Cancer Cell Lines

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Human breast cancer cell lines (BT-20, T-47D, SK-BR-3, MCF-7, MDA-MB-468, MDA-MB-157, BT-474, DU4475, and MDA-MB-231) were all purchased from the American Type Culture Collection, Manassas, VA. An additional MCF-7 cell line derived from Dr. Jose Baselga’s laboratory (Memorial Sloan Kettering Cancer Center, New York, NY) was kindly provided by Dr. Yen-Shen Lu (National Taiwan University Hospital, Taipei, Taiwan) (14 (link)). Cell line authentication by short tandem repeat sequencing was performed to check for cross-contamination. BT-20 cells were grown in Eagle’s Minimum Essential Medium supplemented with 10% fetal bovine serum (FBS). MDA-MB-468 and MDA-MB-231 cells were grown in Leibovitz’s L-15 Medium supplemented with 10% FBS. All cells were maintained at 37°C in a 5% CO₂ humidified atmosphere.
Cisplatin was purchased from Fresenius Kabi, Viman Nagar, India. Phthalic acid was obtained from Sigma-Aldrich, Merck KGaA, Darmstadt, Germany. For specific inhibitors, Y16 was obtained from MedChemExpress, Monmouth Junction, NJ; selisistat and olaparib from Selleck Chemicals, Houston, TX; NF340, Y27632, U73122, and ML7 from ApexBio Technology, Houston, TX.

Liu C.L., Cheng S.P., Chen M.J., Lin C.H., Chen S.N., Kuo Y.H, & Chang Y.C. (2021). Quinolinate Phosphoribosyltransferase Promotes Invasiveness of Breast Cancer Through Myosin Light Chain Phosphorylation. Frontiers in Endocrinology, 11, 621944.

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Isolation and Quantification of Alveolar Type II Cells

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Organoids were retrieved from Matrigel by mechanical dissociation with a P1000, washed 2× with 1 mL PBS and, resuspended with TrypLE and incubated at 37 °C until a single cell suspension was obtained with light pipetting (typically 10 min). Cells were washed 3× with 1 mL of FACS buffer consisting of PBS, 2% BSA and 10 µM Y-27632 (APExBIO, Cat#3008) and then filtered through a 30 µM mesh. Cells were stained for 1 h on ice in FACS buffer with a 1:60 dilution of anti HTII-280 IgM antibody. Cells were washed 3× with 1 mL of FACS buffer before 30 min of staining in FACS buffer with a 1:1000 dilution of anti-mouse IgM-Alexaflour-488 secondary antibody (Jackson Immunoresearch, Cat#715545140) on ice. After 3× washes with 1 mL of FACS buffer cells were suspended in FACS buffer with 1:4000 dilution of DAPI. Live cells (determined by exclusion of DAPI) positive for HTII-280 were identified by 488 emission intensity notably higher than primary only, secondary only controls and absence of 405 (DAPI) emission. All steps were carried out at 4 °C unless otherwise indicated. Cells were pelleted at 500xG for 5 min in swing bucket rotors. FACS was performed on either a Sony MA900 or the ThermoFisher Bigfoot Spectral Cell Sorter and analyzed in FlowJo v10.

Frum T., Hsu P.P., Hein R.F., Conchola A.S., Zhang C.J., Utter O.R., Anand A., Zhang Y., Clark S.G., Glass I., Sexton J.Z, & Spence J.R. (2023). Opposing roles for TGFβ- and BMP-signaling during nascent alveolar differentiation in the developing human lung. NPJ Regenerative Medicine, 8, 48.

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Culture and Expansion of Human Bronchial Epithelial Cells

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Beas-2B immortalized human bronchial epithelial cells were obtained from American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (Corning) with l-glutamine and 4.5 g/l glucose supplemented with 10% fetal bovine serum (VWR) and 1% penicillin/streptomycin (Corning). Deidentified primary human small airway epithelial cells (smAECs; <2 mm diameter) were obtained from the National Jewish Health Biobank and plated onto an irradiated National Institutes of Health (NIH)/3T3 (American Type Culture Collection) fibroblast feeder layer in F-medium containing 1 μM Y-27632 (APEX Bio). Upon visible colony formation (∼7–10 days), smAECs were removed with 0.25% trypsin (Corning), plated on tissue culture dishes double-coated with type I bovine collagen solution (Advanced Biomatrix), and grown to confluence in BronchiaLife Epithelial Medium supplemented with the complete LifeFactors Kit from Lifeline Cell Technology. All cells were maintained in 5% CO₂ at 37 °C.

Gupta A., Sasse S.K., Gruca M.A., Sanford L., Dowell R.D, & Gerber A.N. (2021). Deconvolution of multiplexed transcriptional responses to wood smoke particles defines rapid aryl hydrocarbon receptor signaling dynamics. The Journal of Biological Chemistry, 297(4), 101147.

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