The antibodies used were: anti-SIX3, anti-H3K4me1, anti-H3K4me2, anti-JAG1, anti-GLI1, anti-ZEB2, anti-ANGPTL4 (Abcam, Hong Kong, China), anti-NCOA3 (BD Biosciences, USA), anti-MTA3, anti-MBD2/3, anti-H3pan-ac (Millipore, Billerica, MD, USA), anti-LSD1, anti-HDAC1, anti-HDAC2, anti-RbAp46/48 (Sigma-Aldrich, St Louis, MO, USA), anti-MTA1, anti-MTA2, and anti-WNT1 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA). Human recombinant Wnt1 was purchased from Sigma-Aldrich. Protein A/G Sepharose CL-4B beads were from Amersham Biosciences (Indianapolis, IN, USA) and shRNAs were from GenePharma Co., Ltd (Shanghai, China).
Anti wnt1
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-WNT1 is a laboratory research reagent that targets the WNT1 protein. WNT1 is a signaling protein involved in various cellular processes. Anti-WNT1 can be used to detect and study the WNT1 protein in experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Anti h3k4me1, by Abcam (1 mentions)
Anti h3k4me2, by Abcam (1 mentions)
Anti hdac2, by Merck Group (1 mentions)
Anti hdac1, by Merck Group (1 mentions)
Anti lsd1, by Merck Group (1 mentions)
Anti zeb2, by Abcam (1 mentions)
Anti gli1, by Abcam (1 mentions)
Anti angptl4, by Abcam (1 mentions)
Anti mta3, by Merck Group (1 mentions)
Anti mbd2 3, by Merck Group (1 mentions)
Anti rbap46 48, by Merck Group (1 mentions)
Anti mta1, by Santa Cruz Biotechnology (1 mentions)
Anti mta2, by Santa Cruz Biotechnology (1 mentions)
Protein a g sepharose cl 4b beads, by Cytiva (1 mentions)
Anti jag1, by Abcam (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
Anti c jun, by Santa Cruz Biotechnology (1 mentions)
Anti β catenin, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti mmp7, by Santa Cruz Biotechnology (1 mentions)
4 protocols using anti wnt1
1
Comprehensive Chromatin Regulatory Network
2
Protein Extraction and Western Blotting from Tumor Tissues
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Total protein extraction from tumor tissues using RIPA buffer (Cell Signaling Technology, Inc. Danvers, MA) and western blotting were performed as described previously studies [18 (link), 19 (link)]. The following primary antibodies were used: anti-Wnt-1, anti-β-catenin, anti-c-myc, anti-cyclin D1, anti-MMP-7, anti-axin-2, anti-APC, anti-c-jun, or anti-β-actin antibodies (Santa Cruz Biotechnology, Inc.). The secondary antibodies were corresponding to the type of primary antibody using anti-mouse or anti-rabbit IgG horseradish peroxidase (HRP)-conjugate (1:1,000 dilution, Cell Signaling Technology, Inc.). Protein bands were visualized by the Enhanced Chemiluminescence kit. The quantification of protein bands were measured by FluorChem Imaging Systems (Alpha Innotech). The data were standardized by β-actin.
3
Immunohistochemical Analysis of Wnt1 and CD44 in Gastric Cancer
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Formalin-fixed and paraffin-embedded gastric malignant and nonmalignant tissue sections were incubated with rabbit polyclonal antibody anti-Wnt1 (dilution 1 : 200, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and antibody anti-CD44 (dilution 1 : 200, Santa Cruz Biotechnology) overnight at 4 °C in a humidified container. Detection was determined by non-biotin horseradish peroxidase detection system and DAB substrate (Dako, Carpinteria, CA, USA). Immunostaining was scored by two independent observers. Briefly, five random views of stained sections were chosen and total 100 tumor cells per view were counted for analysis. Expression of Wnt1 or CD44 in less than 10% of tumor cells examined was defined as negative, expression in 10–29% was defined as low expression, 30–60% was defined as moderate expression, and >60% was defined as intense expression.46 (link)
4
Western Blot Analysis of Wnt Signaling
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Cells were lysed using radioimmunoassay precipitation assay (RAPA) lysis buffer with a proteinase inhibitor. The quantity of approximately 40 µg proteins in the lysates were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene di uoride (PVDF) membranes (Millipore, Billerica, MA, USA). The membranes were incubated with anti-WNT1 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), GSK3β (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), β-catenin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and GAPDH (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) for 12 h at 4 °C and then washed three times for 5 min.
Following horseradish peroxidase-conjugated secondary antibodies were incubated. The blot was visualized by an enhanced chemiluminescence kit (Thermo Fisher Scienti c, Inc., Rockford, IL, USA).
Following horseradish peroxidase-conjugated secondary antibodies were incubated. The blot was visualized by an enhanced chemiluminescence kit (Thermo Fisher Scienti c, Inc., Rockford, IL, USA).
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