NSCs that were cultured on coverslips were fixed with 4.5% PFA in PBS, washed with PBS, blocked in 5% donkey serum (Merck Life Science, catalog no. S30-100ML) in PBST (PBS + 0.2% Triton X-100, VWR, catalog no. 0694-1 L), and incubated overnight with primary antibodies (anti‐TCF7L2, Cell Signaling Technology, catalog no. 2569, 1:250 dilution; anti-SOX9, Biotechne, catalog no. AF3075-SP, 1:250 dilution; anti-GFAP, Merck Life Science, catalog no. Hpa056030, 1:250 dilution). The next day, the coverslips were washed and incubated with Alexa Fluor‐conjugated secondary antibodies (donkey anti-rabbit immunoglobulin G [IgG] with Alexa Fluor 488, Invitrogen, catalog no. A‐21206, 1:500 dilution; donkey anti-mouse IgG with Alexa Fluor 594, Invitrogen, catalog no. A‐21203, 1:500 dilution; donkey anti-mouse Author: IgG with Alexa Fluor 555, Invitrogen, catalog no. A-31570, 1:500 dilution; donkey anti-Goat IgG with DyLight 650, Invitrogen, catalog no. SA5-10089, 1:500 dilution). Finally, the coverslips were mounted on microscope slides with Fluoromount-G Mounting Medium with DAPI (Invitrogen, catalog no. 00-4959-52). Images were captured with an Axio Imager Z2 LSM 700 Zeiss confocal microscope.
Anti tcf7l2
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-TCF7L2 is an antibody product that recognizes the Transcription Factor 7 Like 2 (TCF7L2) protein. TCF7L2 is a key transcription factor involved in the Wnt signaling pathway. The antibody can be used to detect and study the expression and localization of TCF7L2 in various cellular and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti gst, by Thermo Fisher Scientific (2 mentions)
Anti ha, by Roche (2 mentions)
Anti flag hrp, by Merck Group (2 mentions)
Anti parp, by Cell Signaling Technology (2 mentions)
Anti gfap, by Merck Group (1 mentions)
Donkey serum, by Merck Group (1 mentions)
Fluoromount g mounting medium with dapi, by Thermo Fisher Scientific (1 mentions)
Dylight 650, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 555, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Anti β catenin, by Cell Signaling Technology (1 mentions)
Anti flag antibody, by Merck Group (1 mentions)
Anti his antibody, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Non immune igg, by Santa Cruz Biotechnology (1 mentions)
Bioruptor ucd 200, by Diagenode (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti p stat3, by Cell Signaling Technology (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
7 protocols using anti tcf7l2
1
Immunofluorescence Staining of Neural Stem Cells
2
Antibody-Based Protein Detection and Enrichment
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Western blots were probed with the following antibodies: Anti-Flag-HRP (Sigma-Aldrich, A8592), Anti-HA (Roche, 11867423001), Anti-His (Cytiva, 27-4710–01), and Anti-GST (Invitrogen, A-5800). Anti-Flag immunoprecipitations were carried out using magnetic beads pre-conjugated with the antibody (Millipore, M8823). Anti-TCF7L2 (Cell Signaling Technology, 2569S) antibodies were also used for ChIP.
3
Western Blot Antibody Validation
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Western blot analysis was performed, as previously described (40 (link)). Anti-Flag antibody was purchased from Sigma-Aldrich (St. Louis, MO). Anti–β-tubulin antibody was purchased from The Developmental Studies Hybridoma Bank. Anti-PARP, anti-caspase3, anti-Tcf7l2, and anti–β-catenin antibodies were purchased from Cell Signaling Technology (Danvers, MA). Anti-His antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-GLUT1 and anti-Zic2 antibodies were purchased from Abcam (Cambridge, MA).
4
ChIP-seq protocol for TCF7L2
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ChIP was performed in triplicate following the instructions provided by the suppliers of the Pierce Agarose ChIP Ki (Thermo). Briefly, MIN6 cells were cross-linked with 1% formaldehyde for 20 min at room temperature and sonicated on ice for 15 cycles of 30 s on and 30 s off (Bioruptor UCD-200; Diagenode, Liège, Belgium). The sonicated chromatin was primarily in the 200 to 1000 bp range, which was successively centrifuged for 15 min and incubated overnight at 4 °C with either Anti-TCF7L2 (Cell Signaling Technology, USA) and non-immune IgG (Santa Cruz Biotechnologies, USA) for negative control at 1 µg of antibody per 106 cells. The precipitated chromatin was eluted at room temperature for 20 min, de-cross-linked by incubation at 65 °C for 5 h in the presence of 200 mM NaCl, extracted with phenol–chloroform, and precipitated.
5
Protein Expression Analysis of Pancreatic Cells
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Cultured islets or Min6 cells were washed in PBS and lysed. PVDF membranes were incubated with anti-TCF7L2 (#2565), anti-actin (#4967), anti-p-AKT (Serine473, #9271), anti-AKT (#9272), anti-p-GSK3β (Ser9 #9336), anti-PARP (#9542), anti-GAPDH (#2118), anti-c-casp3 (#9661), anti-stat3 (#9132), anti-p-stat3 (Tyr705, #9131), anti-PKA C-α ( #5842; all from Cell Signaling, Danvers, MA, USA), anti-β-catenin (ab6302), anti-GLP-1R (ab39072), anti-p-Jak2 (ab68268; all from Abcam), followed by incubation with horseradish-peroxidase-linked IgG peroxidase. The bands were visualized and densities of the bands were analyzed using Tanon ChemImaging Systems (Nanjing, China).
6
Western Blot Analysis of Thalamocortical Proteins
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Protein extracts were obtained from six animals per genotype from at least two litters. The thalamo-habenular regions were dissected from the brains (Fig. S7A,B ) and homogenised in ice-cold RIPA buffer. Protein concentrations were determined using the Bio-Rad protein assay (5000006, Bio-Rad Laboratories). Clarified protein homogenate (50 μg) was loaded on 10% SDS-polyacrylamide gels. Separated proteins were then transferred to Immun-Blot PVDF membranes (1620177, Bio-Rad Laboratories), which were then blotted with anti-TCF7L2 (1:1000; 2569, Cell Signaling Technology), anti-β-actin (1:500; A3854, Sigma-Aldrich) and anti-GAPDH (1:1000; SC-25778, Santa Cruz Biotechnology) antibodies. The staining was visualised with peroxidase substrate for enhanced chemiluminescence (ECL) and 200 μM coumaric acid. Images were captured using the Amersham Imager 600 RGB (General Electric).
7
Western Blot and ChIP Analyses
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Western blots were probed with the following antibodies: Anti-Flag-HRP (Sigma-Aldrich, A8592), Anti-HA (Roche, 11867423001), Anti-His (Cytiva, 27-4710-01), and Anti-GST (Invitrogen, A-5800). Anti-Flag immunoprecipitations were carried out using magnetic beads pre-conjugated with the antibody (Millipore, M8823). Anti-TCF7L2 (Cell Signaling Technology, 2569S) antibodies were also used for ChIP.
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