Phospho β catenins33 s37 t41

Manufactured by Cell Signaling Technology

Phospho-β-cateninS33/S37/T41 is a primary antibody that recognizes β-catenin when phosphorylated at Ser33, Ser37, and Thr41. It is used to detect the phosphorylation of β-catenin at these specific residues.

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Lab products found in correlation

6 protocols using phospho β catenins33 s37 t41

Breast Cancer Cell Lines Protocol

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MDA-MB-231, SKBR3, and MCF7 human BC cell lines were obtained from the American Type Culture Collection (ATCC) (Manassas, VA) and maintained 231 cells (ATCC) was maintained in Dulbecco’s modified Eagle’s medium (DMEM) (4.5 g/L glucose) supplemented with 10% fetal bovine serum, Penicillin (100 µg/ml)/streptomycin (100 µg/mL) antibiotics. Cells were incubated under humidified conditions with 5% CO₂ at 37 °C.
Antibodies against phospho-β-catenin^S552, phospho-β-catenin^S33/S37/T41, β-catenin, E-cadherin, N-cadherin, Vimentin, CD44, phospho-GSK3β^S9, GSK3β, phospho-AKT^T308, AKT, pan-cytokeratin and Actin were purchased from Cell Signaling Technologies, Inc (Boston, MA). An anti-HNA antibody was purchased from Abcam (Cambridge, UK). p16^INK4a was purchased from Abcam.
Cycloheximide (CHX) was purchased from Sigma-Aldrich (St. Louis, MO) and NS1643 was purchased from Alomone Labs (Jerusalem, Israel).

Breuer E.K., Fukushiro-Lopes D., Dalheim A., Burnette M., Zartman J., Kaja S., Wells C., Campo L., Curtis K.J., Romero-Moreno R., Littlepage L.E., Niebur G.L., Hoskins K., Nishimura M.I, & Gentile S. (2019). Potassium channel activity controls breast cancer metastasis by affecting β-catenin signaling. Cell Death & Disease, 10(3), 180.

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Oxidative Stress Pathway Protein Analysis

Cited 2 times

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Dihydroethidium (Cat. # 50-850-563) was purchased from Thermo Fisher Scientific. Paraquat dichloride (Cat # 856177-1G) was purchased from Sigma-Aldrich. The mammalian nuclear isolation kit was purchased from Thermo Fisher Scientific (Cat. #78833). Rabbit polyclonal antibodies against GAPDH (VWR; Cat. # 89348-232), PGK1 (Thermo Fisher; Cat. # PA528612), LRP6 (Cell Signaling; Catalog # 2560), phospho-β-catenin S33/S37/T41 (Cell Signaling; Catalog # 9561), β-catenin (Abcam; Cat # 6302), Phospho-LRP-6 T1479 (Abnova; Cat. # PAB12632) [19 (link)], Lamin A/C (Cell Signaling; Cat. # 2032S), CK1γ3 (Thermo Fisher; Cat # PA5-99838) [20 (link)], Prdx1 (Millipore Sigma; Cat. # HPA007730-100UL), Trx1 (Cell Signaling; Cat. # 2429), Gpx1 (Abcam; Cat. # ab22604), and Prdx-SO₃ (Abcam; Cat. # Ab16830), and mouse monoclonal antibodies against Porin (Invitrogen; Cat. #459500) and Prdx2: PRDX2 (Fisher Scientific; Cat. # LFMA0144) were purchased from commercial sources as indicated. A previously described rabbit polyclonal antibody against Sod1 was obtained from the laboratory of Valeria Culotta (Johns Hopkins University) [16 (link)]. Goat α-rabbit (VWR; Cat # 89138-520) or α-mouse (VWR; Cat # 89138-516) secondary antibodies conjugated to a 680-nm emitting fluorophore were obtained from Biotium through VWR.

Chandrasekharan B., Montllor-Albalate C., Colin A.E., Andersen J.L., Jang Y.C, & Reddi A.R. (2020). Cu/Zn Superoxide Dismutase (Sod1) regulates the canonical Wnt signaling pathway. Biochemical and biophysical research communications, 534, 720-726.

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Regulation of β-Catenin Signaling by SB-216763

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HeLa cells (2 × 10⁸ cells/well) were treated with vehicle or with 0.1, 1 or 10 µM SB-216763 for 30 min and then with 50 nM calyculin at 37 °C. Lysates were prepared as described above, separated by SDS-PAGE, and transferred to PVDF membrane. Immunoblotting was performed using primary and secondary antibodies at the indicated dilutions in TBST: β-catenin (Cell Signaling, Cat. No. 8480, 1:1000) or phospho-β-catenin-S33/S37/T41 (Cell Signaling, Cat. No. 9561, 1:1000) and goat anti-rabbit IgG-HRP (Calbiochem, Cat. No. 12-348, 1:25,000). Blots were developed with chemiluminescence, imaged by film, and quantified as above. To measure GS activity, serum-starved HeLa cells were treated for 60 min with 10 µM Bio, BIM-IX or SB-216763 or DMSO vehicle. Cells were harvested, non-denaturing lysates prepared, and supernatants assayed for GS activity using the GCS activity assay kit (Cat. No. AK0138-100T-96S, Sunlong) by monitoring the oxidation of NADH oxidation to NAD⁺ at 340 nm.

Ferreira R.B., Fu L., Jung Y., Yang J, & Carroll K.S. (2022). Reaction-based fluorogenic probes for detecting protein cysteine oxidation in living cells. Nature Communications, 13, 5522.

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Antibodies for Western Blotting and Immunofluorescence

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The following antibodies were used for western blotting, immunoprecipitations, and immunofluorescence: total β-catenin (1:2,000) (Cell Signaling #9562), phospho-β-catenin S33/S37/T41 (1:1,000) (Cell Signaling #9561), phospho-β-catenin S45 (1:1,000) (Cell Signaling #9564), α-tubulin (1:2,000) (Cell Signaling #2144), PARP (1:1,000) (Cell Signaling #9542), pY99 (1:500) (Santa Cruz sc-7020), HA (1:1,000) (Cell Signaling #3724), Myc (1:1,000) (Cell Signaling #2276), Flag (1:100,000) (Sigma #F3165), GST (1:1,000) (Cytiva 27-4577-01), E-cadherin (1:1,000) (BD #610181), E-cadherin (Proteintech #20874-1-AP), phospho-Lrp6 S1490 (1:1,000) (Cell Signaling #2568), phospho-Tau S202/T205 (1:1,000)(Cell Signaling #30505). Secondary antibodies used for western blotting were anti-rabbit-HRP (1:5,000 or 1:10,000) (Jackson ImmunoResearch #115-035-003), anti-mouse-HRP (1:5,000 or 1:10,000) (Jackson ImmunoResearch #111-035-003), IRDye 800CW goat anti-mouse (1:16,666) (LI-COR Biosciences #926-32210), IRDye 680RD goat anti-mouse (1:33,333) (LI-COR Biosciences #926-68070). Where applicable, western blotting dilutions are denoted in parentheses.

Dutt P., Haider N., Mouaaz S., Podmore L, & Stambolic V. (2024). β-catenin turnover is regulated by Nek10-mediated tyrosine phosphorylation in A549 lung adenocarcinoma cells. Proceedings of the National Academy of Sciences of the United States of America, 121(19), e2300606121.

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Antibodies for Cellular and Molecular Analysis

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Antibodies recognising GFP (S268B), CK1α (SA527), CK1δ (SA609), CK1ε (SA610), and FAM83F (SA103) are available on request from the MRC-PPU reagents website (http://mrcppureagents.dundee.ac.uk). Antibodies for β-actin (#4967), GAPDH (14C10) (#2118), p-LRP6 (S1490) (#2568), LRP6 (C47E12) (#3395), β-catenin (D10A8) (#8480), phospho-β-catenin (S45) (#9564), phospho-β-catenin (S33/S37/T41) (#9561), Na/K ATPase (D4Y7E) (#23565), and Lamin A/C (#2032) were purchased from Cell Signalling Technology. Additional antibodies used were FAM83G (ab121750; Abcam), α-tubulin (MA1-80189; Thermo Fisher Scientific), Active-β-catenin (anti-ABC) clone 8E7 (05-665; END Millipore), mouse anti-GFP clone 7.1 and 13.1 (Roche), mouse-monoclonal anti-HA (H9658; Sigma-Aldrich), and U2AF1 (PA5-28510; Thermo Fisher Scientific). Secondary antibodies used were StarBright Blue 700 Goat anti-Rabbit IgG (12004161; Bio-Rad), StarBright Blue 700 Goat anti-Mouse IgG (12004158; Bio-Rad), IRDye 800CW Donkey anti-Goat IgG (926-32214; LI-COR), IRDye 800CW Goat anti-Rat IgG (926-32219; LI-COR), IRDye 800CW Goat anti-Mouse (926-32210; LI-COR), and IRDye 680LT Goat anti-Rabbit (926-68021; LI-COR).

Dunbar K., Jones R.A., Dingwell K., Macartney T.J., Smith J.C, & Sapkota G.P. (2020). FAM83F regulates canonical Wnt signalling through an interaction with CK1α. Life Science Alliance, 4(2), e202000805.

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Comprehensive Antibody Analysis for Signaling Pathways

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COL5A2 antibody (PA5-14245) was purchased from ThermoFisher Scientific, and COL5A2 antibody (TA313657, OriGene) was used for immunohistochemistry (IHC) analysis. Antibodies for human phospho-p70S6K (Thr389) (#9234), phospho-mTOR (Ser2448) (#2971), phospho-GSK-3α/β (Ser21/9) (#8566), GSK-3α/β (#5676), phospho-Akt (Ser473) (#4060), phospho-Akt (Thr308) (#4056), β-catenin (#8480), phospho-PDK1 (Ser241) (#3061), APC (#2504), AXIN1 (#3323), CK1 (#2655), phospho-β-cateninS33/S37/T41 (#9561), phospho-β-cateninS45 (#9564), non-phospho (active) β-cateninS33/37/T41 (#8814), C-MYC (#9402), DDR1 (#3917) and pY792 DDR1 (#11994) were purchased from Cell Signaling Technology. β-actin antibody (60008-1-Ig) and Cyclin D1 antibody (60186-1-Ig) were purchased from Proteintech Group. E-cadherin antibody (no. 610181) was purchased from BD Transduction Laboratories. VEGF antibody (sc-152) was purchased from Santa Cruz Biotechnology. GAPDH antibody was purchased from Kangwei Century Co. LTD, Beijing, China.

Wang J., Jiang Y.H., Yang P.Y, & Liu F. (2021). Increased Collagen Type V α2 (COL5A2) in Colorectal Cancer is Associated with Poor Prognosis and Tumor Progression. OncoTargets and therapy, 14, 2991-3002.

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