The primary antibodies were rabbit anti-GEFT (Abcam, ab127690), rabbit anti-Snail (Abcam, ab229701), mouse anti-Slug (Abcam, ab51772), mouse anti-Twist (Abcam, ab175430), rabbit anti-Rac1 (Abcam, ab33186), mouse anti-E-cadherin (CST, #14472), rabbit anti-N-cadherin (CST, #13116), rabbit anti-PAK1 (Abcam, ab223849), Cdc42 (Abcam, ab187643), rabbit anti-RhoA (Abcam, ab187027), mouse anti-Zeb1 (Santa Cruz, sc-81428), mouse anti-Zeb2 (Santa Cruz, sc-271984), and mouse anti-β-actin (ZSGB-BIO, China). The secondary antibody was peroxidase-conjugated goat anti-mouse/rabbit IgG (ZB-2305). Cdc42 Activation Assay Biochem Kit™ (cytoskeleton, Cat. # BK034), Rac1 Activation Assay Biochem Kit™ (cytoskeleton, Cat. # BK035) and Rho A Activation Assay Biochem Kit™ (cytoskeleton, Cat. # BK036) were used for analysis of Cdc42, Rac1 and Rho A activation. NSC23766 (Selleck, S8031), ZCL278 (Selleck, S7293), and IPA-3 (Selleck, S7093) inhibitors were acquired commercially.
Zcl278
Manufactured by Selleck Chemicals
Sourced in United States
ZCL278 is a laboratory equipment product designed for scientific research and analysis. It is a precision instrument that serves a core function within the laboratory setting. The technical details and specifications of this product are available upon request.
Automatically generated - may contain errors
Lab products found in correlation
Nsc23766, by Selleck Chemicals (2 mentions)
Mouse anti zeb2, by Santa Cruz Biotechnology (1 mentions)
Cdc42 activation assay biochem kit, by Cytoskeleton (1 mentions)
Rabbit anti pak1, by Abcam (1 mentions)
Mouse anti β actin, by ZSGB-BIO (1 mentions)
Rac1 activation assay biochem kit, by Cytoskeleton (1 mentions)
Rhoa activation assay biochem kit, by Cytoskeleton (1 mentions)
Ipa 3, by Selleck Chemicals (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Selleck Chemicals (1 mentions)
Ccg 1423, by Selleck Chemicals (1 mentions)
Necrox 5, by Enzo Life Sciences (1 mentions)
A375sm, by Korean Cell Line Bank (1 mentions)
Dulbecco s modified, by Thermo Fisher Scientific (1 mentions)
Ml141, by Selleck Chemicals (1 mentions)
Picm0rg50, by Merck Group (1 mentions)
Ham s f 12 nutrient mixture dmem f12, by Thermo Fisher Scientific (1 mentions)
6 well culture dishes, by NEST Biotechnology (1 mentions)
4 protocols using zcl278
1
Investigating EMT Regulators and Signaling
2
Cdc42 Protein Synthesis and Purification
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ZCL278 was purchased from Selleck Chemicals (Houston, TX, USA). Human recombinant Cdc42 protein was purchased from Abcam (Cambridge, MA, USA).
3
Culturing Human Melanoma Cell Lines
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Human melanoma cell lines (A375P, A375SM) were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea). The A375P cell was maintained in DMEM supplemented with 10% (v/v) fetal bovine serum (FBS), 1% (v/v) L-glutamine and penicillin–streptomycin. A375SM was maintained in MEM supplemented with 10% (v/v) FBS and 1% (v/v) L-glutamine and penicillin–streptomycin. All cells were cultured in a 37 °C, 5% CO2 humidified incubator. NecroX-5 (Enzo Life Sciences, Farmingdale, NY, USA), ZCL278, NSC23766, and CCG-1423 (Selleck Chemicals, Houston, TX, USA) were dissolved in dimethyl sulfoxide (DMSO) for each condition and dose. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and DMSO were purchased from Sigma–Aldrich (St. Louis, MO, USA).
4
CDC42 Regulation in Mouse Ovary Culture
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Neonatal females were sacrificed by cervical dislocation on the designated time. Mouse ovaries were separated by microdissection in cold phosphate buffered saline (PBS) under a stereomicroscope (ZSA302, COIC, China) in sterile conditions. The isolated ovaries were cultured on an insert (PICM0RG50, Millipore, USA) in 6-well culture dishes (NEST, China) in 1200 μL Dulbecco’s modified Eagle’s medium/Ham’s F12 nutrient mixture (DMEM/F12) (GIBCO, Life Technologies, USA) plus ITS (1:100, Sigma, USA) and Penicillin-Streptomycin Solution at 37 °C, 5% CO2 and saturated humidity. Ovaries were cultured for 2 or 5 days to assess the role of CDC42, in either medium alone or medium supplemented with ML141 (5uM, Selleck, China) or ZCL278 (50uM, Selleck, China). ML141 is a selective reversible CDC42 inhibitor. ZCL278 is a selective CDC42 inhibitor.
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