The cells’ protein was collected by RIPA lysis buffer (Byotime, P0045), then the BCA protein assay kit (Biosharp, BL521A) normalizes the samples. The cell extracts were boiled with Sample Loading Buffer (Biosharp, BL529A) for 5 min. After separated by 10% SDS-PAGE gels and transferred to PVDF membrane (Millipore, Germany), the membranes were blocked in 5%BSA and incubated with primary antibodies at 4°C overnight. The primary antibodies are listed as follows: anti-PLAU (1:2,000, Proteintech, 17968-1-AP), anti-E-Cadherin (1:5,000, Proteintech, 20874-1-AP), anti-N-Cadherin (1:2,000, Proteintech, 22018-1-AP), anti-TWIST1 (1:1,000, Proteintech, 25465-1-AP), anti-SNAI1 (1:1,000, Proteintech, 13099-1-AP). Then the membranes were incubated with secondary antibody (Biosharp, BL003A) and the proteins were visualized by ECL enhanced chemiluminescence substrate (Millipore).
Anti snai1
Manufactured by Proteintech
Anti-SNAI1 is a laboratory reagent used to detect and quantify the SNAI1 protein, a transcription factor involved in epithelial-mesenchymal transition. This antibody can be used in various applications such as Western blotting, immunohistochemistry, and immunocytochemistry to study the expression and localization of SNAI1 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti e cadherin, by Proteintech (2 mentions)
Anti n cadherin, by Proteintech (2 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Bca protein assay kit, by Biosharp (1 mentions)
Bl003a, by Biosharp (1 mentions)
Anti plau, by Proteintech (1 mentions)
Anti twist1, by Proteintech (1 mentions)
Anti mmp2, by Proteintech (1 mentions)
Anti mmp9, by Proteintech (1 mentions)
Mouse anti β actin, by Merck Group (1 mentions)
Rabbit anti mouse igg, by Abcam (1 mentions)
Anti vimentin, by Proteintech (1 mentions)
2 protocols using anti snai1
1
Western Blot Analysis of EMT Markers
2
Western Blot Analysis of Protein Markers
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Western blotting was performed according to a previous protocol (2 (link)). Rabbit polyclonal anti-PLCD3, anti-MMP2, anti-MMP9, anti-Snai1, anti-E-cadherin, anti-vimentin, anti-N-cadherin and anti-Flot2 were purchased from ProteinTech. Mouse anti-β-actin was purchased from Sigma. Rabbit anti-mouse IgG was purchased from Abcam (1:40,000; Cambridge, MA, USA). Quantification of signal intensity [integral optical density (IOD)] was performed with Gel-Pro Analyzer software (version 4.0). Expression changes in protein levels were assessed by IOD, and the intensity was normalised to the β-actin signal. This experiment was independently repeated three times.
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