Our Abpromise guarantee covers the use of ab12793 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Blocking - Blocking peptide for Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody - ChIP Grade (ab5095)
- First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
- If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
- Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
- Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
- Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Information available upon request.
ab12793 RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2 0.1 µg/ml was mixed with GST tagged recombinant proteins in vitro in binding buffer for 1 hour at RT. Glutathione coated magnetic beads were then mixed into the buffer and removed after another hour incubation. The GST tagged recombinant proteins and any bound peptide was eluted by incubating with free glutathione. The peptides analyzed by dotblot using primary antibody ab5095(1/3000) and an undiluted conjugated HRP-Goat anti-rabbit secondary antibody.
This experiment was conducted with ab12795, ab18488 and an another peptide as a negative control.
This peptide was used as a positive control for ab5095 (anti S2p-CTD)
Sample: Human Recombinant protein (ab12793 peptide)
Loading amount: 0.25 µg
Specification: ab12793 peptide
Gel Running Conditions: Reduced Denaturing (16% tris-tricine (Schagger et al 2006))
Blocking step: BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
Other product details
Incubation time: 1 hour(s) and 0 minute(s) · Temperature: 23°C
Secondary antibody: HRP conjugated Goat anti-rabbit polyclonal, at a 1/20,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"