Ovarian and breast cancer cells were collected in RIPA buffer (Thermo Scientific; Rockford, IL) containing 1% Halt Proteinase Inhibitor Cocktail (Thermo Scientific; Rockford, IL). An equal amount of protein (40 µg/lane) was loaded onto 10% SDS-PAGE gels and transferred onto nitrocellulose membranes. The membranes were blocked with 5% non-fat milk for 1 h and incubated with primary antibodies against KLF4 (Cell Signaling), GAPDH (Sigma; St. Louis, MO), vimentin, E-cadherin, or snail2 (Cell Signaling).
Snail2
Manufactured by Cell Signaling Technology
Sourced in United States
Snail2 is a lab equipment product offered by Cell Signaling Technology. It is a protein that plays a role in cellular processes, but a detailed description of its core function cannot be provided without additional interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
E cadherin, by Cell Signaling Technology (5 mentions)
Vimentin, by Cell Signaling Technology (5 mentions)
Ripa buffer, by Thermo Fisher Scientific (3 mentions)
Halt proteinase inhibitor cocktail, by Thermo Fisher Scientific (3 mentions)
Gapdh, by Merck Group (2 mentions)
β catenin, by Cell Signaling Technology (2 mentions)
Gapdh, by Santa Cruz Biotechnology (2 mentions)
Eif5a2, by Abcam (1 mentions)
Cytokeratin 7, by Abcam (1 mentions)
Smad2, by Cell Signaling Technology (1 mentions)
P smad2, by Cell Signaling Technology (1 mentions)
Pdcd4, by Cell Signaling Technology (1 mentions)
Aicar, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Compound c, by Merck Group (1 mentions)
Secondary antibody conjugated to horseradish peroxidase, by Thermo Fisher Scientific (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Haematoxylin, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dapi dimethyl sulfoxide dmso, by Merck Group (1 mentions)
6 protocols using snail2
1
Protein Expression Analysis of Cancer Cells
2
Western Blot Analysis of Ovarian Cancer
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Western blot (WB) was performed as described previously [30 (link)]. Briefly, ovarian cancer cells were collected in RIPA buffer (Thermo Scientific; Rockford, IL) containing 1% Halt Proteinase Inhibitor Cocktail (Thermo Scientific; Rockford, IL). Equal amounts of protein (100 μg/lane) were loaded onto 10% SDS-PAGE gels and transferred onto nitrocellulose membranes. The membranes were blocked with 5% nonfat milk for 1 h and incubated with primary antibodies against EIF5A2, Cytokeratin-7(Abcam), GAPDH (Santa Cruz; St. Louis, MO), Vimentin, Ecadherin, β-catenin, snail2, SMAD2 or p-SMAD2 (Cell Signaling).
3
Ovarian Cancer Protein Expression Analysis
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Ovarian cancer cells were collected in RIPA buffer (Thermo Scientific; Rockford, IL) containing 1% Halt Proteinase Inhibitor Cocktail (Thermo Scientific; Rockford, IL). An equal amount of protein (40 μg/lane) was loaded onto 10% SDS-PAGE gels and transferred onto nitrocellulose membranes. The membranes were blocked with 5% nonfat milk for 1 h and incubated with primary antibodies against PDCD4 (Cell Signaling), GAPDH (Sigma; St. Louis, MO), vimentin, E-cadherin, or snail2 (Cell Signaling), cytokeratin-7.
4
Signaling Pathway Modulation in Cancer
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AICAR, compound C, eosin, haematoxylin, 4′,6‐diamidino‐2‐phenylindole (DAPI), dimethyl sulfoxide (DMSO) and crystal violet were obtained from Sigma‐Aldrich (St. Louis, MO, USA). Simvastatin was purchased from LKT Laboratories, Inc (St. Paul, MN, USA), and pitavastatin calcium was purchased from HL Genomics (Gyeonggi‐Do, Republic of Korea). LY294002 and antibodies against p‐Akt (Ser437), Akt, p‐AMPK (Tyr172), AMPK, p‐FOXO3a (Ser253, Ser413), FOXO3a, PUMA, Caspase‐3, Caspase‐9, PARP, E‐cadherin, N‐cadherin, vimentin, β‐catenin, ZEB1, Snail2 and Lamin B1 were purchased from Cell Signaling Technology (Beverly, MA, USA), and GAPDH was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). A secondary antibody conjugated to horseradish peroxidase was procured from Thermo Fisher Scientific (Rockford, IL, USA).
5
Comprehensive Protein Expression Analysis
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Western blot procedure was performed according to the method as previously described (14 (link)). The primary antibodies E-cadherin (BD Biosciences, catalog no: 610181 RRID:AB_397581), N-cadherin (BD Biosciences, catalog no: 610920 RRID:AB_398236), Vimentin (BD Biosciences, catalog no: 550513 RRID:AB _393716), iNOS (BD Biosciences, catalog 610332, RRID:AB_397722), β-actin (Sigma-Aldrich, catalog no: MAB1501 clone C4; catalog no: A2066 RRID:AB_476693), Cleaved caspase-9 (Cell Signaling Technology, catalog no: 9502 RRID:AB_2068621); (Cell Signaling Technology, catalog no: 7237 RRID:AB_10895832), Fibronectin [Cell Signaling Technology, catalog no: 26836 Fibronectin/FN1 (E5H6X) Rabbit mAb], Gata4 (Cell Signaling Technology, catalog no: 36966 RRID:AB_2799108), Lamin A/C (Cell Signaling Technology, catalog no: 4777, RRID:AB_10545756), Snail2 (Cell Signaling Technology, catalog no: 9585, RRID:AB_2239535), Twist1 (Cell Signaling Technology, catalog no: 69366, RRID:AB_2891135), Zeb2 [Santa Cruz Biotechnology, catalog no: sc-271984 (E-11)], CD45 (Miltenyi Biotec, catalog no: 130–115–938, RRID:AB_2751284), EpCam (Invitrogen, catalog no: MA5–13917 RRID:AB_11001308); (Novus Biologicals, catalog no: NBP2–27107), CRB2 (ThermoFisher Scientific, catalog no: PA5–25628, RRID:AB_2543128).
6
EMT Marker Expression in SKOV3 Cells
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To detect the expression of EMT-associated marker genes, KLF4-expressing and control SKOV3 cells were fixed for 10 min using 4% PFA, washed three times with 0.1% Tween20 in PBS (PBST), and incubated with blocking buffer (5% normal goat serum, 3% bovine serum albumin, and 0.1% Triton-X 100 in PBS) for 1 h. The primary antibodies to E-cadherin, snail2, and vimentin (1∶200 dilution, Cell Signaling, Danvers, MA), were incubated with fixed cells overnight. After rinsing three times for 5 min with PBST, Alexa 488 or 594 conjugated goat anti-rabbit (1∶200 dilution, Life Technologies) antibodies were added for 1 h at room temperature. Cell nuclei were counterstained with DAPI (Vector Laboratories, Inc.; Burlingame, CA). Images were taken using a Nikon inverted fluorescence microscope.
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