Anti lef1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-LEF1 is a laboratory antibody product used for the detection and analysis of the LEF1 (Lymphoid Enhancer Binding Factor 1) protein. LEF1 is a transcription factor that plays a role in the Wnt signaling pathway and is involved in cellular processes such as cell proliferation and differentiation. The Anti-LEF1 antibody can be used in various applications, including Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of the LEF1 protein in biological samples.

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Lab products found in correlation

Anti tg2, by Cell Signaling Technology (1 mentions) Mab645, by R&D Systems (1 mentions) Anti wnt5a, by R&D Systems (1 mentions) Anti lamin a c sc 6215, by Santa Cruz Biotechnology (1 mentions) Anti phospho β catenin, by Cell Signaling Technology (1 mentions) Ab32572, by Abcam (1 mentions) Anti ar, by Santa Cruz Biotechnology (1 mentions) Tyramide superboost kit, by Thermo Fisher Scientific (1 mentions) Abc kit, by Vector Laboratories (1 mentions) Anti β catenin, by Abcam (1 mentions) Phosphatase, by Santa Cruz Biotechnology (1 mentions) Hematoxylin counterstain, by Thermo Fisher Scientific (1 mentions) Anti c met, by Cell Signaling Technology (1 mentions) Dab chromogen, by Vector Laboratories (1 mentions) Anti yap1, by Santa Cruz Biotechnology (1 mentions) Anti cd8, by Agilent Technologies (1 mentions) Vulcan fast red chromogen, by Biocare Medical (1 mentions) Anti cd163, by Abcam (1 mentions) Ripa buffer, by Merck Group (1 mentions) Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)

5 protocols using anti lef1

Investigating Wnt Signaling Pathway

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Anti LEF1 (sc374522, Santa Cruz); anti β−catenin (ab32572, AbCam); anti phospho β−catenin (9561, Cell Signaling); anti TG2 (3557, Cell signaling); anti TG2 (sc71632, Santa Cruz); anti Lamin A/C (sc-6215, Santa Cruz); anti Tubulin (T-4026, Sigma); anti Wnt5a (MAB645, R&D); anti Wnt10b (PA5-20530, Invitrogen).

Rossin F., Costa R., Bordi M., D’Eletto M., Occhigrossi L., Farrace M.G., Barlev N., Ciccosanti F., Muccioli S., Chieregato L., Szabo I., Fimia G.M., Piacentini M, & Leanza L. (2021). Transglutaminase Type 2 regulates the Wnt/β-catenin pathway in vertebrates. Cell Death & Disease, 12(3), 249.

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Immunohistochemical Analysis of Genital Tissues

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Immunohistochemistry was performed according to standard procedures using anti-BrdU (G3G4, DSHB, Cat# AB 2314035, RRID: AB_2618097), anti-AR (Santa Cruz Biotechnology, Cat# sc-816, RRID: AB_1563391), anti-β-catenin (Abcam, Cat# ab32572, RRID: AB_725966), anti-frizzled-6 (Santa Cruz Biotechnology, Cat# sc-393791, RRID: AB_2736833), and anti-LEF1 (Santa Cruz Biotechnology, Cat# sc-374522, RRID: AB_10986008), and AR and LEF1 antibodies were detected using Tyramide SuperBoost Kits (Invitrogen, B40922) and ABC Kit (Vector Laboratories, PK-6100), respectively, according to the manufacturers’ protocols. Cell proliferation and programmed cell death analyses were performed using BrdU and LysoTracker, respectively, following the previously published methods [10 (link)]. A Leica DM5500 Confocal Microscope (Leica microsystems Inc, Buffalo Grove, IL) was used for imaging. Sections of GTs (Sample size, n = 5) from three different litters were selected.

Wang S., Lawless J, & Zheng Z. (2020). Prenatal low-dose methyltestosterone, but not dihydrotestosterone, treatment induces penile formation in female mice and guinea pigs. Biology of Reproduction, 102(6), 1248-1260.

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Cellular Fractionation and Immunohistochemistry

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Cell lysates for Western blots were made in RIPA buffer (Sigma) supplemented with protease (Roche) and phosphatase (Santa Cruz Biotechnology) inhibitor cocktails. We used the NE-PER® Nuclear and Cytoplasmic extraction reagents (Pierce Biotechnology) for cellular fractionation. For IHC, after deparaffinization and rehydration, tissue sections were antigen-retrieved at 95°C for 30 minutes. Immunostaining with anti-c-MET (Cell Signaling Technology) was performed using a streptavidin–biotin, horseradish peroxidase and DAB chromogen (Vector Labs). IHC with anti-YAP1 (Santa Cruz Biotechnology) and anti-LEF1 (Santa Cruz Biotechnology) was performed using alkaline phosphatase, vulcan fast red chromogen (Biocare Medical), and hematoxylin counterstain (Thermo Scientific). Immunostaning with anti-CD8 (Dako) and anti-CD163 (Abcam) was visualized by TRITC- and FITC-labeled secondary antibodies, respectively, and nuclei were counterstained by DAPI.

Hugo W., Shi H., Sun L., Piva M., Song C., Kong X., Moriceau G., Hong A., Dahlman K.B., Johnson D.B., Sosman J.A., Ribas A, & Lo R.S. (2015). Non-genomic and Immune Evolution of Melanoma Acquiring MAPKi Resistance. Cell, 162(6), 1271-1285.

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Protein Expression Analysis in Cell Lines

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Cells were harvested in cell lysis buffer (Beyotime, China), and the protein content was quantified by BCA methods. The protein was separated onSDS-PAGE and transferred onto a nitrocellulose membrane. The membrane was incubated with the primary anti-EZH2, anti-β-actin and anti-β-catenin (Abcam, UK), anti-c-Myc, anti-CyclinD1, anti-TCF-1, anti-LEF-1 (Santa Cruz, USA), and active anti-β-Catenin (Millipore, USA) and secondary horseradish peroxidase-conjugated antibody. β-actin was used as an internal control.

Wang H., Meng Y., Cui Q., Qin F., Yang H., Chen Y., Cheng Y., Shi J, & Guo Y. (2016). MiR-101 Targets the EZH2/Wnt/β-Catenin the Pathway to Promote the Osteogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells. Scientific Reports, 6, 36988.

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Western Blot Analysis of Intestinal Proteins

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Freshly isolated crypts, enteroids, or sorted intestinal epithelial cells were suspended in ice-cold, radioimmunoprecipitation assay buffer (Cell Signaling, Danvers, MA) containing Halt Protease inhibitor (ThermoFisher Scientific) and lysed at 4°C by supersonication. Total lysate protein was loaded on 10% sodium dodecyl sulfate–polyacrylamide gels for electrophoresis, membrane transfer, and immunoblotting. Anti-active β-catenin (1:2000 dilution, 05-665; Millipore), anti-Lef1 (1:500 dilution, sc-28687; Santa Cruz, Dallas, TX), and anti-Cftr 3G11 (1:2000 dilution, provided by CFTR Folding Consortium, Cystic Fibrosis Foundation Therapeutics) were used as primary antibodies. Anti–β-actin (1:2000 dilution, sc-130656; Santa Cruz) or anti–glyceraldehyde-3-phosphate dehydrogenase (1:2000 dilution, sc-25778; Santa Cruz) were used as loading controls. Densitometry was performed using Image Lab Software (version 5.2.1; BioRad).

Strubberg A.M., Liu J., Walker N.M., Stefanski C.D., MacLeod R.J., Magness S.T, & Clarke L.L. (2017). Cftr Modulates Wnt/β-Catenin Signaling and Stem Cell Proliferation in Murine Intestine. Cellular and Molecular Gastroenterology and Hepatology, 5(3), 253-271.

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