Ad-GFP or Ad-BMP9 infected subconfluent MEFs cells. At 48 hours after infection, cells were cross-linked and subjected to ChIP analysis as previously described [56 (link),57 (link),58 (link)]. Smad1/5/8 antibody (Santa Cruz Biotechnology) or control IgG was used to pull down the protein-DNA complexes. The presence of RUNX1promoter sequence was detected by using two pairs of primers corresponding to mouse RUNX1 promoter region.
Smad1 5 8 antibody
Manufactured by Santa Cruz Biotechnology
The Smad1/5/8 antibody is a laboratory tool used for the detection and analysis of the Smad1, Smad5, and Smad8 proteins in various biological samples. These proteins are members of the Smad family, which are key mediators of the transforming growth factor-beta (TGF-β) signaling pathway. The Smad1/5/8 antibody can be utilized in techniques such as Western blotting, immunohistochemistry, and immunofluorescence to study the expression and localization of these proteins in cells and tissues.
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Lab products found in correlation
2 protocols using smad1 5 8 antibody
1
ChIP Analysis of Smad1/5/8 Binding
2
Embryonic Transcription Factor ChIP-seq
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Majority of ChIP-seq datasets were obtained from NCBI's Gene Expression Omnibus (see Key Resources Table ). For newly generated datasets, ChIP-seq was performed as previously described (Chiu et al., 2014) at the appropriate developmental stage. The antibodies and conditions for these datasets:
• 30 ug of published VegT antibody (Sudou et al., 2012) per 2000-3000 embryos
• 30 ug of published Mix1 antibody (Sudou et al., 2012) per 2000-3000 embryos
• 30 ug of published Sia1 antibody (Sudou et al., 2012) per 2000-3000 embryos
• 4ug of Sox7 rabbit polyclonal peptide antibody (Genscript) per 100 embryos; the peptide antibody was designed against a region in the Sox7 transactivation domain in the Cterminus with the sequence QVSQASDIQPSETS • 3.5 ug of Ventx2 rabbit polyclonal antibody per 100 embryos; the antibody was generated by Covance, Inc., using a GST fusion to Ventx2.2 amino acids 2-153, upstream of the homeodomain.
• 2.5 ug of Smad1/5/8 antibody (Santa Cruz Biotechnology sc-6031x) per 100 embryos Libraries were generated using NEXTflex ChIP-seq (Bioo Scientific, Cat# NOVA-5143-01) kit, quality tested using an Agilent Bioanalyzer 2100, quantified using KAPA qPCR, and sequenced using Illumina sequencers at the UC Irvine Genomics High Throughput Facility.
ATAC-seq was available from BioRxiv: doi.org/10.1101/2020.02.26.966168 (Bright et al.,2020) (link).
• 30 ug of published VegT antibody (Sudou et al., 2012) per 2000-3000 embryos
• 30 ug of published Mix1 antibody (Sudou et al., 2012) per 2000-3000 embryos
• 30 ug of published Sia1 antibody (Sudou et al., 2012) per 2000-3000 embryos
• 4ug of Sox7 rabbit polyclonal peptide antibody (Genscript) per 100 embryos; the peptide antibody was designed against a region in the Sox7 transactivation domain in the Cterminus with the sequence QVSQASDIQPSETS • 3.5 ug of Ventx2 rabbit polyclonal antibody per 100 embryos; the antibody was generated by Covance, Inc., using a GST fusion to Ventx2.2 amino acids 2-153, upstream of the homeodomain.
• 2.5 ug of Smad1/5/8 antibody (Santa Cruz Biotechnology sc-6031x) per 100 embryos Libraries were generated using NEXTflex ChIP-seq (Bioo Scientific, Cat# NOVA-5143-01) kit, quality tested using an Agilent Bioanalyzer 2100, quantified using KAPA qPCR, and sequenced using Illumina sequencers at the UC Irvine Genomics High Throughput Facility.
ATAC-seq was available from BioRxiv: doi.org/10.1101/2020.02.26.966168 (Bright et al.,2020) (link).
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