Total protein was extracted by cell lysate (BB-3209, Bestbio, Shanghai, China), separated by SDS-PAGE, and then transferred onto a polyvinylidene fluoride membrane. After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c-myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3βSer9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA). The membrane was then incubated with the secondary antibody goat anti-rabbit antibody to immunoglobulin G (IgG) (A21020, Abbkine, USA, 1:1,000) for incubation for 1 h at 37°C. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was regarded as the loading control. The expression of target protein is equal to the gray value of target protein bands divided by the gray value of GAPDH.
Ab75629
Manufactured by Abcam
Sourced in United States, United Kingdom
Ab75629 is an antibody product manufactured by Abcam. It is designed for use in laboratory research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab131097, by Abcam (1 mentions)
Ab15148, by Abcam (1 mentions)
Ab93926, by Abcam (1 mentions)
Ab39688, by Abcam (1 mentions)
Ab6302, by Abcam (1 mentions)
Ab137321, by Abcam (1 mentions)
Ab82846, by Abcam (1 mentions)
Ab83042, by Abcam (1 mentions)
Ab61758, by Abcam (1 mentions)
A21020, by Abbkine (1 mentions)
Bb 3209, by BestBio (1 mentions)
Chemiluminescence detection kit, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Sds loading buffer, by Beyotime (1 mentions)
Ab9869, by Abcam (1 mentions)
2 protocols using ab75629
1
Western Blot Analysis of Wnt Pathway
2
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The total cell extracts were processed by denaturing proteins with SDS loading buffer (Beyotime, China). Equivalent amounts of protein were separated by 12% SDS-polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes (Millipore Corporation, MA, and U.S.A.) and then blocked for 30 min in Tris-buffered saline with Tween 20 (TBST) and 5% non-fat milk. The blots were incubated at 4°C overnight in a series of antibodies diluted in TBST buffer: BCL11A (Clone: EPR 14943-44, Abcam, London, U.K.) and MDM2 primary antibodies (1:5,000), baculoviral IAP repeat containing 3 (BIRC3) antibody (Clone: E 40, 1:1000 dilution, Abcam), β-2-microglobulin (B2M) antibody (Clone: EP2978Y, 1:5000 dilution, Abcam), STMN1 antibody (Clone: EP1573, 1:50000 dilution, Abcam), SNAI2 antibody (ab75629, 1:50000 dilution, Abcam), haem oxygenase 1 (HMOX1) antibody (Clone: HO-1-1, 1:1000 dilution, Abcam), and protein phosphatase 1 regulatory inhibitor subunit 15A (PPP1R15A) antibody (ab 9869, 1:1000 dilution, Abcam). After washing with TBST, the membranes were incubated in secondary antibodies (1:4,000; KPL, MD, USA) and then analyzed by a chemiluminescence detection kit (Thermo Scientific, Inc., NJ, USA).
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