PHF7Flag-HA was co-expressed with GFP, Gata4Myc, Hand2Myc or Mef2cMyc, or SMARCD3Flag-HA was co-expressed with PHF3xTy1 or GFP in HEK293 cells for 72 h. Experiments involving crosslinking were adapted from Zlatic et al.37 , whereby cells were treated with 200 μM dithiobis(succinimid ylpropionate) (DSP) or dimethylsulfoxide (DMSO) for 4 h at 4 °C and quenched with 25 mM Tris for 15 min. Pre-cleared lysates were incubated with Flag magnetic beads (Sigma, M8823) overnight. The Flag epitope tag was eluted using 0.5 mg ml−1 free 3× Flag peptide (Sigma). The final elution and input obtained before immunoprecipitation were analysed by western blotting using an anti-Myc (Invitrogen), anti-Ty1 (Diagenode) or rabbit anti-Flag antibody (Sigma).
Anti ty1
Manufactured by Diagenode
Anti-Ty1 is a laboratory reagent used in the detection and analysis of Ty1 proteins. It provides a reliable and specific method for identifying and quantifying Ty1 proteins in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Flag magnetic beads, by Merck Group (1 mentions)
Flag peptide, by Merck Group (1 mentions)
Anti myc, by Thermo Fisher Scientific (1 mentions)
Rabbit anti flag antibody, by Merck Group (1 mentions)
Monoclonal anti gfp, by Roche (1 mentions)
Enhanced chemi luminescence (ecl), by GE Healthcare (1 mentions)
2 protocols using anti ty1
1
Protein Interaction Profiling in HEK293 Cells
2
Immunodetection of Protein Markers in Malaria Parasites
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The probes were collected from ~7–8% synchronized schizont cultures. The parasite material was suspended in SDS sample buffer and incubated at 90 °C for 5 minutes. Proteins were separated on 10–15% SDS-PAGE and subsequently transferred onto a nitrocellulose membrane. Immunoblots were performed using standard procedures and developed by chemiluminescence using ECL (GE Healthcare). Antibodies used in immunodetection were rabbit monoclonal anti-TY1 (Diagenode) and monoclonal anti-GFP (Roche). anti-TY1 was used 1:1000, anti-GFP antibody 1:1000 and anti-Aldolase 1:5000 diluted in phosphate-buffered saline (PBS) with 5% w/v skim milk. Secondary antibodies were sheep anti-rabbit IgG horseradish peroxidase (Sigma) and sheep anti-mouse IgG horseradish peroxidase (Roche) used 1:5000 diluted in PBS with 5% w/v skim milk.
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