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ROCK Inhibitor (Y-27632)

ROCK Inhibitor (Y-27632)

(RUO)
ROCK Inhibitor (Y-27632)
ROCK inhibitor, Y-27632, improves the recovery of human embryonic stem cells (hESC) after fluorescence activated cell sorting. H9 hESC (WiCell, Madison, WI) were grown in mTESR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277), dissociated with BD™ Accutase™ Cell Detachment Solution (Cat. No. 561527), and sorted on a BD FACSAria™ II cell sorter based upon cell-surface expression of SSEA-3, TRA-1-81 and SSEA-1 using the BD Stemflow™ Human Pluripotent Stem Cell Sorting and Analysis Kit, (Cat. No. 560461). ROCK Inhibitor was either omitted or added for 24 hours post-sort. Bright-field images of the hESC at days 1 and 3 after sorting and culturing without or with ROCK Inhibitor are shown. Day 1, no treatment (Top left panel). Day 3, no treatment (Top right panel). Day 1, 10 µM Y-27632 treatment (Bottom left panel). Day 3, 10 µM Y-27632 treatment (Bottom right panel). For details please see Emre et al., 2010.
ROCK inhibitor, Y-27632, improves the recovery of human embryonic stem cells (hESC) after fluorescence activated cell sorting. H9 hESC (WiCell, Madison, WI) were grown in mTESR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277), dissociated with BD™ Accutase™ Cell Detachment Solution (Cat. No. 561527), and sorted on a BD FACSAria™ II cell sorter based upon cell-surface expression of SSEA-3, TRA-1-81 and SSEA-1 using the BD Stemflow™ Human Pluripotent Stem Cell Sorting and Analysis Kit, (Cat. No. 560461). ROCK Inhibitor was either omitted or added for 24 hours post-sort. Bright-field images of the hESC at days 1 and 3 after sorting and culturing without or with ROCK Inhibitor are shown. Day 1, no treatment (Top left panel). Day 3, no treatment (Top right panel). Day 1, 10 µM Y-27632 treatment (Bottom left panel). Day 3, 10 µM Y-27632 treatment (Bottom right panel). For details please see Emre et al., 2010.
Product Details
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BD™
Y-27632 dihydrochloride
Cell culture (Tested During Development)
AB_2869435
RUO


Preparation And Storage

Store at room temperature, desiccated and protected from prolonged exposure to light. Store aliquots of the reconstituted material at -20°C, protected from prolonged exposure to light, and avoid repeated freeze-thaw cycles.

Recommended Assay Procedures

For a working solution, dilute to 10 mM in sterile deionized water. [The molar mass of Y-27632 is 338.3 g/mol.]

Small aliquots may be stored at -20°C, avoiding repeated freeze-thaw cycles.

Use at 10 µM in culture medium. Since applications vary, each investigator should titrate the reagent to obtain optimal results.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Avoid contact with skin and eyes.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. mTESR™1 is a trademark of StemCell Technologies.
  5. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
562822 Rev. 1
Antibody Details
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The ROCK inhibitor, Y-27632, is a selective inhibitor of the p160-Rho-associated coiled kinase (ROCK) and is a factor that enhances embryonic stem cell survival upon single cell dissociation. In addition to increasing cell recovery after dissociation, Y-27632 has been used for various other applications in stem cell research including cryopreservation, sorting, reprogramming, transplantation, and differentiation.

562822 Rev. 1
Citations & References
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Development References (5)

  1. Braam SR, Nauw R, Ward-van Oostwaard D, et al. Inhibition of ROCK improves survival of human embryonic stem cell-derived cardiomyocytes after dissociation. Ann N Y Acad Sci. 2010; 1188:52-57. (Methodology: Cell culture). View Reference
  2. Emre N, Vidal JG, Elia J, et al. The ROCK inhibitor Y-27632 improves recovery of human embryonic stem cells after fluorescence-activated cell sorting with multiple cell surface markers. PLoS ONE. 2010; 5(8):e12148. (Methodology: Cell culture). View Reference
  3. Li X, Meng G, Krawetz R, Liu S, et al. The ROCK inhibitor Y-27632 enhances the survival rate of human embryonic stem cells following cryopreservation. Stem Cells Dev. 2008; 17(6):1079-1085. (Methodology: Cell culture). View Reference
  4. Narumiya S, Ishizaki T, Uehata M. Use and properties of ROCK-specific inhibitor Y-27632. Methods Enzymol. 2000; 325:273-284. (Biology). View Reference
  5. Watanabe K, Ueno M, Kamiya D. A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol. 2007; 25(6):681-686. (Methodology: Cell culture). View Reference
View All (5) View Less
562822 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.