After treatment, total proteins were extracted from cells using radioimmunoprecipitation assay lysis buffer (RIPA) with 1 mM phenylmethanesulfony fluoride (PMSF) (both from Beyotime Biotechnology, Shanghai, China) and quantified using a bicinchoninic acid (BCA) assay. Twenty micrograms of proteins were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels (12%) and transferred onto a PVDF membrane. Subsequently, the membrane was blocked with 5% non-fat milk in TBST solution for 2 h at room temperature (RT). After incubation with the primary antibodies against Wnt-4 (sc-5214), Frizzled-2 (sc-68327; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), β-catenin (9582s), GSK-3β (9315; Cell Signaling Technology, Inc., Beverly, MA, USA), cyclin D1 (BS6532), Col-II (BS1071; Bioworld Technology, Natong, China) overnight at 4°C, and the HRP-conjugated secondary antibodies (ZB-2301; Zhongshan Golden Bridge Biotechnology Co., Ltd.) for 1 h at RT, the blots were detected using a Bio-Rad Chemi Doc XRS+ (Bio-Rad), and β-actin was used as the control.
Frizzled 2
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Frizzled-2 is a protein that functions as a receptor for Wnt signaling pathways. It is a member of the Frizzled protein family and plays a role in cellular processes such as cell fate determination, cell migration, and embryonic development.
Automatically generated - may contain errors
Lab products found in correlation
Chemidoc xr, by Bio-Rad (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
Zb 2301, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Anti human gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti mouse igg hrp conjugate, by Promega (1 mentions)
2 protocols using frizzled 2
1
Protein Expression Analysis Protocol
2
Wnt Signaling Pathway Antibodies
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Human Phospho-β-Catenin (Ser33/37/Thr41) Antibody (1:1000, #9561, CST, Boston, MA,USA), Human anti-wif-1 antibody (1:1000, #5652, CST), Human anti-DNMT1(1:1000, #5032, CST), Frizzled 5 (1:1000, #5266, CST), Frizzled 2 (1:200, sc-74,019, Santa Cruz, CA, USA), Human anti-GAPDH (1:200, sc-47,724, Santa Cruz), anti-mouse IgG HRP conjugate (1:8000, W402B, Promega, Madison, WI, USA), and anti-Rabbit IgG HRP conjugate (1:2000, #7074P2, CST). Indirubin was obtained from Aladdin (#I132661, Shanghai, China) and dissolved in dimethyl sulfoxide (DMSO).
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