CRC cells (4 × 105) were treated with Obatoclax (0, 100, 200 μM) for 24 h, or treated with Obatoclax for 22 h and then combined with MG132 (20 μM) for an additional 2 h. After drug treatment, cells were lysed, subjected to SDS-PAGE, blot transfer, incubation with primary and secondary antibodies, exposure to enhanced chemiluminescence (ECL) reagent, and final visualization of the antigen-of-interest using the LAS3000 system (Fujifilm; Tokyo, JPN) as stated previously [45 (link),46 (link)]. Densitometry analysis of immunoblot signals was accomplished by ImageJ software (National Institute of Health, Bethesda, MD, USA). Anti-survivin (#ab469) and anti-TCF4 (#ab76151) antibodies were bought from Abcam (Cambridge, GBR). Anti-β-catenin (sc-7963) and anti-c-MYC (sc-40) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-LEF1 (GTX129186) and anti-GAPDH (GTX627408) were obtained from GeneTex (Hsinchu, TWN), and anti-β-tubulin (#T2200) was bought from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against cyclin D1 (#2926), HA-tag (#3724), and cleaved forms of caspases-3 (#9664), -8 (#9496), -9 (#9501), and PARP (#9541) were all purchased from Cell signaling Technology (Boston, MA, USA).
Anti β catenin sc 7963
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-β-catenin (sc-7963) is a primary antibody product offered by Santa Cruz Biotechnology. It is designed to detect β-catenin, a key component of the Wnt signaling pathway and an important regulator of cell-cell adhesion and gene transcription.
Automatically generated - may contain errors
Lab products found in correlation
Anti survivin ab469, by Abcam (1 mentions)
Anti c myc sc 40, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh gtx627408, by GeneTex (1 mentions)
Las 3000 system, by Fujifilm (1 mentions)
Ab18197, by Abcam (1 mentions)
Ab98023, by Abcam (1 mentions)
Ab8805, by Abcam (1 mentions)
Bca protein assay kit, by Bioss Antibodies (1 mentions)
Sc 516102 sc 2357, by Santa Cruz Biotechnology (1 mentions)
Sc 8418, by Santa Cruz Biotechnology (1 mentions)
Rpmi 1640 medium, by Yuanye Bio-Technology (1 mentions)
Anti flag 20543 1 ap, by Proteintech (1 mentions)
Anti axin2, by Cell Signaling Technology (1 mentions)
Anti β actin ac026, by ABclonal (1 mentions)
Sr3029, by MedChemExpress (1 mentions)
Anti gfp 66002 1 ig, by Proteintech (1 mentions)
Anti igg 30000 0 ap, by Proteintech (1 mentions)
Anti v5, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh, by Proteintech (1 mentions)
Anti ki67, by BioLegend (1 mentions)
10 protocols using anti β catenin sc 7963
1
Immunoblot Analysis of Apoptosis Regulators
2
Protosappanin B Anti-Cancer Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protosappanin B was purchased from Shanghai Yuanye Bio-Technology Co. Ltd. (B21622-20 mg, Shanghai, China). Reagents, signaling pathway agonists and antibodies were purchased from the following companies: RPMI-1640 medium, DMEM medium (Shanghai Yuanye Bio-technology, Shanghai, China); Streptavidin Peroxidase immunohistochemistry kit (GS4961, GS4962), cell counting kit-8 (QN1293), Annexin V-FITC/PI double staining apoptosis detection kit (SNM530), and ECL western blotting detection kit (QN1155) (Beijing BioRab Technology Co. Ltd., Beijing, China); BCA protein assay kit (C05-02001), western blocking buffer (5% BSA, C05-06002) (Beijing Biosynthesis Biotechnology Co., Ltd, Beijing, China); IGF-1 (K002504P), AZD2858 (A126819), and TPA (bs-1545R) (Shanghai Hengfei Biotechnology Co. Ltd. Shanghai, China); Anti-GOLPH3 (ab98023), Anti-p-AKT (ab8805), Anti-AKT (ab38449), and anti-PCNA (proliferating cell nuclear antigen) (ab18197) (Abcam, Cambridge, UK); Anti-p-p70S6K (ribosomal protein S6 kinase, 70 kDa) (sc-8416), Anti-p70S6K (sc-8418), Anti-β-catenin (sc-7963), Anti-p-ERK1/2(extracellular signal-regulated kinase 1 and 2) (sc-81492), Anti-ERK1/2 (sc-514302), Anti-GAPDH (sc-365062), and secondary antibodies (sc-516102, sc-2357) (Santa Cruz Biotechnology, Inc., Dallas, TX, USA).
3
Antibody Characterization for Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-p53 antibody (SC-126), anti-Caspase-3 (SC-7148), anti-APE1 (SC-55498), and anti-β-catenin (SC-7963) were purchased from Santa Cruz. Anti-CHK1 antibody (#2360) was obtained from CST Biotechnology. Anti-Vinculin antibody (MAB3574) and active-β-catenin (MAB-05665) were obtained from Millipore. Anti-Polβ (ab26343) was obtained from Abcam. Anti-FEN1 (70185) was purchased from Gentex. Anti-TBB5(AM103a), anti-GAPDH (0063), and anti-β-Actin (M0001) were obtained from Abgent.
4
Wnt/β-catenin Pathway Regulation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant human RSPO1 protein (rRSPO1) was purchased from Novo protein (CX83; Shanghai, China), and recombinant Wnt3A protein (rWnt3A) was purchased from R&D Systems (5036-WN-010; Minneapolis, MN, USA). SR3029 was purchased from MedChemExpress (HY-100011; MCE, Monmouth Junction, NJ, USA). MG132 (133407–82-6; Saint Louis, Missouri, USA) was purchased from Sigma–Aldrich. The following primary antibodies were used: anti-CK1δ (sc-55553), anti-Ub (sc-271289) and anti-β-catenin (sc-7963), which were purchased from Santa Cruz Biotechnology; anti-V5 (#13202), anti-LRP6 (#2560), anti-p-LRP6 (Ser1490) (#2568), anti-Axin1 (#2087) and anti-Axin2 (#2151), which were purchased from Cell Signaling Technology; anti-GFP (66002–1-Ig), anti-Flag (20543–1-AP), anti-IgG (30000–0-AP) and anti-GAPDH (600004–1-Ig), which were purchased from Proteintech; anti-CK1ε (ab270997) was purchased from Abcam; and anti-β-actin (AC026), which was purchased from ABclonal.
5
Immunohistochemical Analysis of Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As previously described,24 (link) formalin-fixed tumors were embedded with paraffin and sectioned, after which immunocytochemistry and H&E staining were performed. Immunocytochemistry was performed using the following primary antibodies: anti-Ki-67 (350501; BioLegend), anti-active β-catenin (8814; Cell Signaling), and anti-β-catenin (sc-7963; Santa Cruz).
6
Immunohistochemical Analysis of Mandibular Bone
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry staining was performed on 4-μm-thick coronal sections prepared from 6 mandibles of KO or control mice using a DAB substrate kit (Vector Laboratories, Burlingame, CA, USA) following the manufacturer’s instruction. The primary antibodies used for immunohistochemistry are: anti-phospho-ERK (9101, 1:200, Cell Signaling, Danvers, MA, USA), anti-Lef1 (C12A5, 1:200, Cell signaling, Danver, MA, USA), anti-β-catenin (sc-7963, Santa Cruz), anti-Sox2 (ab97959, Abcam, Cambridge, MA, USA), and anti-phospho-SMAD1/5 (9516, Cell Signaling). Methyl green was used for counterstaining.
7
Cell Lysis and Western Blot Protocol
8
Immunoblot Analysis of β-Catenin Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested by centrifugation and rinsed twice with PBS. Total proteins were harvested from cells, separated on 10% SDS-PAGE gels and then subjected to immunoblot analyses. Primary antibodies against β-catenin (~90 kDa) and β-actin were purchased from Santa Cruz Biotechnology, Inc. (anti-β-catenin, sc-7963; 1:200; anti-β-actin, sc-130301; 1:10,000). The secondary antibody, goat anti-mouse IgG-horseradish peroxidase, was purchased from Santa Cruz Biotechnology, Inc. (sc-2005; 1:5,000). Antibodies bound to the blots were detected using an enhanced chemiluminescence system (Pierce Biotechnology, Inc., Rockford, IL, USA). The mean optical densities (ODs) of β-catenin protein bands were normalized against the OD of the β-actin band from the same treatment. Quantity One software (Bio-Rad Laboratories, Hercules, CA, USA) was used to quantify the OD levels. Immunoblotting experiments were repeated at least 3 times.
9
Western Blot, Immunofluorescence, and Immunohistochemistry Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies anti-human proteins used for Western blot, immunofluorescence and immunohistochemistry were the mouse monoclonal anti-Nodal (ab55676, Abcam, Cambridge, UK), anti-MAP2 (MS-249-R7, ThermoFisher Scientific), anti-PARP1 (sc-8007, Santa Cruz Biotechnology, Dallas, TX, USA), anti-CD44v6 (33-6700, ThermoFisher Scientific), anti-CD31 (M0823, Dako, Agilent Technologies, Santa Clara, CA, USA), anti-Glypican-2 (sc-393824, Santa Cruz Biotechnology), anti-Nucleolin (39-6400, ThermoFisher Scientific), anti-β-catenin (sc-7963, Santa Cruz Biotechnology), anti-LRP6 (sc-25317, Santa Cruz Biotechnology), anti-p53 (sc-126, Santa Cruz Biotechnology), and the rabbit polyclonal anti-N-Cadherin (04-1126, Millipore), anti-Ki-67 (ab833, Abcam), anti-P-Akt and anti-Akt (9271 and 9272, Cell Signaling Technology Inc., Danvers, MA, USA), anti-CD114 (PA5-28988, ThermoFisher Scientific), anti-PES1 (HPA066670, Sigma, St Louis, MO, USA), anti-NPM1 (HPA011384, Sigma), anti-Osteopontin (AB1870, Millipore).
10
Immunofluorescence Analysis of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells grown on coverslips were treated with verteporfin as described above. Cells were fixed in 4% paraformaldehyde were permeabilized with 0.15% Triton X-100 (Sigma) in phosphate-buffered saline or in methanol and were incubated with the following antibodies: anti-YAP (sc-271134, dilution 1:25), anti-β-catenin (sc-7963, dilution 1:50), and anti-ROCK2 (sc-398,519, dilution 1:50) that were all purchased from Santa Cruz Biotechnologies; and anti-N-cadherin (BD Transduction Labs, 610921, dilution 1:100). Anti-mouse FITC (Thermo Scientific, #31569, dilution 1:100) or anti-goat IgG NL493 (FITC equivalent R&D, #NL003, dilution 1:50) were used as secondary antibodies. Nuclei were counterstained with Hoechst 33256 (Sigma). Images were acquired using a Nikon ECLIPSE 90i microscope and were then analyzed with NIS-Elements software (Nikon).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!