Log In Sign up
The largest database of trusted experimental protocols

Ctnnb1

Manufactured by BD
Visit Supplier
Sourced in United States

CTNNB1 is a lab equipment product that functions as a key component in the Wnt signaling pathway. It plays a crucial role in cell-cell adhesion and the regulation of gene transcription.

Automatically generated - may contain errors

Lab products found in correlation

Bca kit, by Thermo Fisher Scientific (3 mentions) Normal mouse igg, by Merck Group (2 mentions) Tcf7l2, by Cell Signaling Technology (1 mentions) Tcf7l1, by Cell Signaling Technology (1 mentions) H3k27ac, by Abcam (1 mentions) Axin1, by Cell Signaling Technology (1 mentions) Sirt1, by Cell Signaling Technology (1 mentions) Criterion tgx precast gel, by Bio-Rad (1 mentions) Ppm1a, by Abcam (1 mentions) Ctnnb1, by Merck Group (1 mentions) Vps35, by GeneTex (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Axioskop 2 fluorescent microscope, by Zeiss (1 mentions) Sc 816, by Santa Cruz Biotechnology (1 mentions) Lsm 880 airyscan, by Zeiss (1 mentions) Dab substrate, by Vector Laboratories (1 mentions) Cyp2e1, by Abcam (1 mentions) Aurora b, by Abcam (1 mentions) Abc elite kit, by Vector Laboratories (1 mentions)

9 protocols using ctnnb1

1

Chromatin Immunoprecipitation in Embryonic Neural Tissues

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
For each experiment, chromatin was isolated from 20 cranial neural folds dissected from HH8-9 embryos or 10 anterior neural plate border regions from HH5 embryos. Immunoprecipitation was performed as described [33 (link)], using the H3K27Ac (Abcam, #ab177178), TFAP2A (DSHB #3B5), LEF1 (Millipore, #17–604), CTNNB1 (BD Biosciences, #610154), TCF7 (Cell Signaling, #C63D9), TCF7L1 (Cell Signaling, #D15G11) and TCF7L2 (Cell Signaling, #C48H11) antibodies and normal mouse IgGs (Millipore, #17–604) as controls. In WNT dominant negative assays, HH4 embryos were electroporated with a WNT dominant negative construct [16 (link)] and at stages HH8-9, cranial neural folds were dissected and processed as described above. The primers used for qPCR quantification are described in S5 Table.
Azambuja A.P, & Simoes-Costa M. (2021). A regulatory sub-circuit downstream of Wnt signaling controls developmental transitions in neural crest formation. PLoS Genetics, 17(1), e1009296.
+ Open protocol
+ Expand
2

Western Blot Analysis of Diverse Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were generated with PBS/1% NP40 buffer supplemented with protease inhibitor cocktail (Sigma Cat. No. S8820). Protein sample loading buffer was added to cell lysates and proteins were separated on SDS-PAGE (BioRad Criterion TGX Precast Gel). The following primary antibodies were used for immunoblotting at the indicated dilutions: Cell Signaling Technology: AXIN1 (2087 and 2074; 1:1000), BAP1 (13271; 1:1000), BCL2L2 (2724; 1:1000), LRP6 (2560; 1:1000), PTEN (9552; 1:1000), RICTOR (9476; 1:1000), SIRT1 (2493; 1:1000), TBK1 (3504; 1:1000), TLE3 (4681; 1:1000), LKB1 (3050; 1:1000), SUFU (2520; 1:1000), LRP5 (5731; 1:1000). Bethyl Laboratories: BAP1 (A302-243A-T; 1:1000), RICTOR (A300-459A; 1:1000), SIRT1 (A300-688A; 1:1000), TOP1 (A302-589A and 302-590A; 1:1000), VPS35 (A304-727A; 1:1000). Abcam: TBK1 (40676; 1:1000), PPM1A (14824; 1:1000), SUFU (52913; 1:1000). Santa Cruz Biotechnologies: LRP6 (sc-25317; 1:1000), TLE3 (sc-9124; 1:1000), LKB1 (sc-374334; 1:1000), LRP5 (sc-390267; 1:1000). LS BioScience: BCL2L2 (LS-C382259-100; 1:1000), PPM1A (LS-C169090-100; 1:1000). Sigma-Aldrich: CTNNB1 (C2206; 1:1000). Invitrogen: PTEN (44-1064; 1:1000). BD Biosciences: CTNNB1 (610153; 1:1000). Genetex: VPS35 (GTC108058; 1:1000).
Tuladhar R., Yeu Y., Tyler Piazza J., Tan Z., Rene Clemenceau J., Wu X., Barrett Q., Herbert J., Mathews D.H., Kim J., Hyun Hwang T, & Lum L. (2019). CRISPR-Cas9-based mutagenesis frequently provokes on-target mRNA misregulation. Nature Communications, 10, 4056.
+ Open protocol
+ Expand
3

Chromatin Immunoprecipitation for Wnt Signaling

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
For each experiment, chromatin was isolated from 16 cranial neural folds dissected from HH8-9 embryos in Ringer’s solution. Immunoprecipitation was performed as described (Simões-Costa et al., 2014 (link)) using the Lef1 (Millipore, #17–604) and Ctnnb1 (BD Biosciences, #610154) antibodies and normal mouse IgGs (Millipore, #17–604) as controls. In Wnt dominant negative assays, HH5 embryos were electroporated with a Wnt dominant negative construct (García-Castro et al., 2002 (link)); at stages HH8-9, cranial neural folds were dissected. Chromatin isolation and immunoprecipitation were performed as previously described (García-Castro et al., 2002 (link)).
Bhattacharya D., Rothstein M., Azambuja A.P, & Simoes-Costa M. (2018). Control of neural crest multipotency by Wnt signaling and the Lin28/let-7 axis. eLife, 7, e40556.
+ Open protocol
+ Expand
4

Immunoblotting of Cell Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in RIPA+ buffer and protein concentrations were analyzed by the BCA kit (Thermo Scientific). Fifteen micrograms of total proteins was resolved on 10% SDS-PAGE gel, transferred to nitrocellulose membranes, and blotted for SMAD2/3 (1:1,000; rabbit IgG, Cell Signaling Technology #8685), CTNNB1 (1:1,000; mouse IgG, BD Biosciences 610154), GAPDH (1:2,000; rabbit IgG, Santa Cruz Biotechnology SC25778), TP53 (1:1,000; rabbit IgG, Santa Cruz SC6243). The secondary antibodies were purchased from Jackson ImmunoResearch.
Liu Z., Hui Y., Shi L., Chen Z., Xu X., Chi L., Fan B., Fang Y., Liu Y., Ma L., Wang Y., Xiao L., Zhang Q., Jin G., Liu L, & Zhang X. (2016). Efficient CRISPR/Cas9-Mediated Versatile, Predictable, and Donor-Free Gene Knockout in Human Pluripotent Stem Cells. Stem Cell Reports, 7(3), 496-507.
+ Open protocol
+ Expand
5

Immunohistochemical Analysis of Testicular and Epididymal Tissues

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Freshly dissected testicular/epididymal tissues were fixed in Bouin’s fixative (H&E, IF) or 4% paraformaldehyde (TUNEL) overnight at 4 °C, and processed for paraffin embedding following routine procedures. IF and TUNEL assays were performed as described previously [11 (link)]. The antibodies and dilutions used were: 1:100 for CUL4B (#20882-1-AP, Proteintech Group, Chicago, IL, USA) and CUL4A (#IHC-00145, Bethyl Laboratories, Montgomery, TX, USA); 1:200 for AR (#sc-816, Santa Cruz Biotechnology, SCBT, Dallas, TX, USA) and SCP3 (#sc-74569, SCBT); 1:500 for VASA (#8761, Cell Signaling Technology, CST, Danvers, MA, USA), CTNNB1 (#610154, BD Biosciences), pS6S235/236 (#2211, CST), pS6S240/244 (#2215, CST), and CLDN11 (#36–4500, ThermoFisher, Waltham, MA, USA); 1:1000 for Alexa 594 goat anti-rabbit, Alexa 488 goat anti-mouse and Alexa 488-conjugated lectin PNA (ThermoFisher). IF images were captured under a Zeiss AxioSkop 2 fluorescent microscope. Confocal microscopy was imaged on Zeiss LSM 880 Airyscan at the Washington University Center for Cellular Imaging.
Yin Y., Zhu L., Li Q., Zhou P, & Ma L. (2021). Cullin4 E3 Ubiquitin Ligases Regulate Male Gonocyte Migration, Proliferation and Blood-Testis Barrier Homeostasis. Cells, 10(10), 2732.
+ Open protocol
+ Expand
6

Immunostaining of Paraffin-Embedded Tissues

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissue samples were fixed in 4% paraformaldehyde (PFA) and embedded in paraffin. In some cases, frozen sections were made. Immunohistochemistry was performed as previously described 31 (link). For immunofluorescence staining, transfected cells were fixed in 4% paraformaldehyde (PFA) at 37C or 15min, followed by blocking with 5% BSA/0.25% Triton in PBS for 1h. Primary antibodies used: Cyp2e1 (Abcam, ab28146), Ctnnb1 (BD Transduction Laboratories #610154), Aurora B (Abcam ab2254), alpha-Tubulin (CST #3873). For immunohistochemistry, detection was performed with the Elite ABC Kit and DAB Substrate (Vector Laboratories), followed by Hematoxylin Solution counterstaining (Sigma). Images were taken by an Olympus IX83 microscope. H&E staining and trichrome staining were performed by the UTSW Histo Pathology Core Facility or the UTSW Tissue Procurement Service. H&E slides were interpreted by P.G., a clinical pathologist with expertise in human liver cancer.
Zhang S., Nguyen L.H., Zhou K., Tu H.C., Sehgal A., Nassour I., Li L., Gopal P., Goodman J., Singal A.G., Yopp A., Zhang Y., Siegwart D.J, & Zhu H. (2017). Knockdown of Anillin Actin Binding Protein Blocks Cytokinesis in Hepatocytes and Reduces Liver Tumor Development in Mice without Affecting Regeneration. Gastroenterology, 154(5), 1421-1434.
+ Open protocol
+ Expand
7

Quantitative Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blotting was performed as previously described 24 . Briefly, 30-60 μg proteins were resolved on 10-12% SDS-PAGE and transferred onto a nitrocellulose membrane (Bio-Rad). Membranes were blocked (milk 5%) then incubated in primary and secondary antibodies. Band intensity was determined and quantified using an Odyssey IR scanner (Li-Cor Biosciences). βactin (1:15000; mouse monoclonal; Sigma #A5316) was used as internal control. The following antibodies were used: Ctnnb1 (1:2000, mouse monoclonal; BD Transduction Laboratories #610153); and goat anti-mouse (IRDye 680 nm, 1:10,000; Li-Cor Biosciences). Samples from different groups were processed on the same blots. Data are expressed as percent relative to controls.
Gapp K., Jawaid A., Sarkies P., Bohacek J., Pelczar P., Prados J., Farinelli L., Miska E, & Mansuy I.M. (2014). Implication of sperm RNAs in transgenerational inheritance of the effects of early trauma in mice. Nature neuroscience, 17(5), 667-669.
+ Open protocol
+ Expand
8

Chromatin Immunoprecipitation (ChIP) Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Crosslinking was performed in 1% paraformaldehyde (15 min, room temperature) and stopped with 0.125 M glycine (5 min). Cells were lysed in PBS containing phosphatase inhibitor mix (PI, Sigma-Aldrich) and 1 mM PMSF, and the pellet was resuspended in 1 ml of ChIP sonication buffer (50 mM HEPES [pH 7.9], 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, 0.1% SDS) also containing PI and PMSF. DNA was sheared by sonication (10 rounds of [10 × 0.5 s on, 0.6 s off, 27% amplitude]) and a sample was used for determining fragment size. DNA (50 μg) was then used for two rounds of immunoprecipitation (round 1: FOXO3a [Santa Cruz, sc-11351]; round 2: CTNNB1 [BD Transduction Laboratories, 610154]), protein was digested using proteinase K, and the resulting sample was used for PCR analysis as described above with primers as listed in Table S2.
McClelland Descalzo D.L., Satoorian T.S., Walker L.M., Sparks N.R., Pulyanina P.Y, & zur Nieden N.I. (2016). Glucose-Induced Oxidative Stress Reduces Proliferation in Embryonic Stem Cells via FOXO3A/β-Catenin-Dependent Transcription of p21cip1. Stem Cell Reports, 7(1), 55-68.
+ Open protocol
+ Expand
9

Comprehensive Antibody Panel for Cell-Cell Adhesion

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse anti-β-catenin (CTNNB1, BD transduction laboratories, #610154), rabbit anti-α-catenin (CTNNA1, Sigma, #C2081), mouse anti-E-cadherin (CDH1, BD transduction laboratories, #610182), mouse anti-phospho-Tyr 4G10 (Millipore, #05-1050), rabbit anti-β-catenin phospho-Tyr142 (Abcam, #ab27798), anti-GM130 (BD transduction laboratories, #610823), rabbit anti-ARVCF (Bethyl Laboratories, #A303-310A), rabbit anti-CDH4 (Sigma, #HPA015613), rabbit anti-ERBB2IP (Sigma, #HPA048606), rabbit anti-PKP4 (Abcam, #ab128638), rabbit anti-PLEKHA6 (Sigma, #HPA028152), rabbit anti-PPP6C (Abcam, #ab70634), rabbit anti-PPP6R3 (Abcam, #ab72034), rabbit anti-HA (Cell signaling technologies, #3724), rabbit anti-TJP2 (ZO-2; Cell signaling technologies, #2847), mouse anti-JUP (Invitrogen, #12-8500), mouse anti-DSG2 (Sigma, #SAB4200466), mouse anti-p-cadherin (CDH3, BD biosciences, #610227), rabbit anti-DSG3(Abcam, #ab128927), mouse anti-PHB (NeoMarkers, #MS-261-PO), Rabbit anti-ITGB4 (Cell signaling technologies, #4707), mouse anti-Integrin β1 (BD biosciences, #552828). A polyclonal antibody against the peptide “NH2-RRLRPGSIETYEQEK-Cys” was generated to detect DUSP23 (Pacific Immunology) and validated using DUSP23 over-expression and knockdown lysates.
Gallegos L.L., Ng M.R., Sowa M.E., Selfors L.M., White A., Zervantonakis I.K., Singh P., Dhakal S., Harper J.W, & Brugge J.S. (2016). A protein interaction map for cell-cell adhesion regulators identifies DUSP23 as a novel phosphatase for β-catenin. Scientific Reports, 6, 27114.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!