ChIP assays were performed as previously described (Gazin et al., 2007 (link)) using the following antibodies: ZNF304 (described above), KAP1 (Bethyl Laboratories), SETDB1 (Millipore, Billerica, MA), DNMT1, DNMT3A, and DNMT3B (all from Imgenex, San Deigo, CA), cJUN (Millipore), H3K27me3 (Cell Signaling Technology), H3K9me3 (Millipore), H3K4me3 (Abcam), EZH2 (Millipore) and BMI1 (Abcam). The CDX1 antibody (Chan et al., 2009 (link)) was kindly provided by Walter Bodmer (University of Oxford, UK). ChIP products were analyzed by qRT-PCR (see Supplementary file 1 for primers). Samples were quantified as percentage of input, and then normalized to an irrelevant region in the genome (∼3.2 kb upstream from the transcription start site of GCLC). Fold enrichment was calculated by setting the IgG control IP sample to a value of 1.
Setdb1
Manufactured by Merck Group
SETDB1 is a protein-coding gene that acts as a histone methyltransferase, responsible for the addition of methyl groups to specific lysine residues on histone proteins. This enzymatic activity plays a role in the regulation of gene expression and chromatin structure.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (3 mentions)
Protease inhibitor cocktail, by Roche (3 mentions)
Dnmt1, by Novus Biologicals (2 mentions)
Znf304, by Fortis Life Sciences (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
H3k9me3, by Merck Group (1 mentions)
Dnmt3a, by Novus Biologicals (1 mentions)
Dnmt3b, by Novus Biologicals (1 mentions)
C jun, by Merck Group (1 mentions)
H3k27me3, by Cell Signaling Technology (1 mentions)
H3k4me3, by Abcam (1 mentions)
Dnmt3b, by Abcam (1 mentions)
Bach1, by Santa Cruz Biotechnology (1 mentions)
α tubulin, by Merck Group (1 mentions)
Cyclin b1, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
C myc, by Santa Cruz Biotechnology (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
6 protocols using setdb1
1
Chromatin Immunoprecipitation Assay Protocol
2
ChIP Assay Protocol with Antibodies
3
Western Blot Analysis of Cell Markers
4
Western Blotting Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed as previously described [33 (link), 34 (link)]. Briefly, cell lysis was performed with the RIPA buffer protein extraction reagent (Pierce, Rockford, IL, USA) containing a protease inhibitor cocktail (Roche, USA). The proteins were resolved by 10% SDS-PAGE followed by transfer to polyvinylidene fluoride (PVDF) membranes (Sigma-Aldrich). Next, the membranes were blocked using 5% bovine serum albumin (BSA) and incubated with primary antibodies at 4 °C overnight. Appropriate secondary antibodies were later added and then visualized by using an ECL chemiluminescence kit. The primary antibodies used are listed as follows: SETDB1 (HPA018142, Sigma-Aldrich), cleaved caspase 3 (9661, Cell signaling technology), cleaved caspase 8 (9748, Cell signaling technology), slug (9585, Cell signaling technology), vimentin (5741, Cell signaling technology), E-cadherin (14,472, Cell signaling technology), mTOR (2983, Cell signaling technology), p-mTOR (5536, Cell signaling technology), AKT (4685, Cell signaling technology), p-AKT (4060, Cell signaling technology), CSF-1 (3155, Cell signaling technology), β-actin (3700, Cell signaling technology).
5
Western Blotting for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed as previously described [29, 30] . Brie y, cell lysis was performed with the RIPA buffer protein extraction reagent (Pierce, Rockford, IL, USA) containing a protease inhibitor cocktail (Roche, USA). The proteins were resolved by 10% SDS-PAGE followed by transfer to polyvinylidene fluoride (PVDF) membranes (Sigma-Aldrich). Next, the membranes were blocked using 5% bovine serum albumin (BSA) and incubated with primary antibodies at 4°C overnight. Appropriate secondary antibodies were later added and then visualizedby using an ECL chemiluminescence kit. The primary antibodies used are listed as follows: SETDB1 (HPA018142, Sigma-Aldrich), cleaved caspase
6
Western Blot for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed as previously described [33, 34] . Brie y, cell lysis was performed with the RIPA buffer protein extraction reagent (Pierce, Rockford, IL, USA) containing a protease inhibitor cocktail (Roche, USA). The proteins were resolved by 10% SDS-PAGE followed by transfer to polyvinylidene fluoride (PVDF) membranes (Sigma-Aldrich). Next, the membranes were blocked using 5% bovine serum albumin (BSA) and incubated with primary antibodies at 4°C overnight. Appropriate secondary antibodies were later added and then visualized by using an ECL chemiluminescence kit. The primary antibodies used are listed as follows: SETDB1 (HPA018142, Sigma-Aldrich), cleaved caspase 3 (9661, Cell signaling
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!