We performed ChIP using the Simple ChIP ® Enzymatic Chromatin IP Kit (ChIP) Kit (Cell Signaling, #9003, USA) according to the manufacturer's instructions. The first step is to lyse the tumor cells and crosslink proteins to DNA. Subsequently, the lysates of first step were sonicated into 300 bp to 600 bp fragments and immunoprecipitated with anti-TEAD1 antibody (Abcam, UK). In addition, we used normal rabbit immunoglobulin G (IgG) as a negative control. Then, we performed reverse crosslinking and DNA purification. Finally, the precipitated DNA was analyzed by qRT-PCR via SYBR-Green PCR Master Mix (Vazyme, Nanjing, China). Primers used for SNHG16 promoter regions are shown in the Supplementary Table 1 .
Anti tead1 antibody
Manufactured by Abcam
Sourced in United Kingdom
Anti-TEAD1 antibody is a laboratory reagent used for the detection and analysis of the TEAD1 protein in biological samples. TEAD1 is a transcription factor involved in cellular processes. This antibody can be used in various experimental techniques, such as western blotting, immunohistochemistry, and immunoprecipitation, to study the expression and localization of TEAD1 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Simplechip enzymatic chromatin ip kit chip kit, by Cell Signaling Technology (2 mentions)
Sybr green pcr master mix, by Vazyme (2 mentions)
Radioimmunoprecipitation assay lysis buffer, by Merck Group (1 mentions)
Goat anti rabbit igg h l hrp, by Abcam (1 mentions)
Anti e cadherin antibody, by Abcam (1 mentions)
Rabbit anti n cadherin antibody, by Abcam (1 mentions)
Rabbit anti yap1 antibody, by Abcam (1 mentions)
Rabbit anti ki67 antibody, by Abcam (1 mentions)
Anti gapdh antibody, by Abcam (1 mentions)
3 protocols using anti tead1 antibody
1
ChIP Assay of TEAD1 Binding to SNHG16
2
Western Blot Analysis of YAP1 Signaling
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Cells were lysed in radioimmunoprecipitation assay lysis buffer (Sigma–Aldrich, MO, USA). After electrophoresis on a 12% or 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel, proteins were transferred onto polyvinylidene difluoride membranes. Subsequently, the membranes were blocked with 5% non-fat milk and incubated overnight at 4°C with the following primary antibodies: Rabbit Anti-YAP1 antibody (1:5000 dilution; Abcam), Rabbit Anti-YAP1 (phosphorylated S127) (pYAP1) antibody (1:50000 dilution; Abcam), Rabbit Anti-Ki67 antibody (1:5,000 dilution; Abcam), Rabbit Anti-TEAD-1 antibody (1:50000 dilution; Abcam), Rabbit Anti-E-cadherin antibody (1:50000 dilution; Abcam), Rabbit Anti-N-cadherin antibody (1:1000 dilution, Abcam), and Rabbit Anti-glyceraldehyde-3-phosphate dehydrogenase (Anti-GAPDH) antibody (1:10000 dilution, Abcam). The HRP-conjugated secondary antibody Goat Anti-Rabbit IgG H&L (HRP) (1:20000 dilution, Abcam, USA) was added and incubated at room temperature for 1 h. Signals were visualized after chemiluminescence reaction with horseradish peroxidase substrate. The relative protein expressions of YAP1, pYAP1, Ki67, Tead, EMT markers (E-Cadherin and N-Cadherin) in different cells were normalized to the GAPDH concentration, and the quantitative analysis was performed using the Image J software (National Institutes of Health (NIH), USA).
3
TEAD1 Chromatin Immunoprecipitation Protocol
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We performed ChIP using the SimpleChIP® Enzymatic Chromatin IP Kit (ChIP) Kit (Cell Signaling, #9003, USA), as per the instruction of manufacturer. Put simply, after the cells were crosslinked and lysed, the cells become lysates. Subsequently, the lysates were sonicated into 300 bp to 600 bp fragments and immunoprecipitated by anti-TEAD1 antibody (Abcam). Besides, we used Normal rabbit immunoglobulin G (IgG) as a negative control. Then, we performed the reverse crosslinking and DNA puri cation. Finally, the precipitated DNA was analyzed by qRT-PCR via the SYBR-Green PCR Master Mix (Vazyme, Nanjing, China). Primers used for SNHG16 promoter regions were shown in supplementary material.
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