Western blot Abreview for RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2

Western blot

Review text: 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

Primary antibody (in addition to 'RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2')
Primary antibody: Abcam primary antibody: Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody - ChIP Grade (ab5095)
Dilution: 1/3000

Secondary antibody
Name: Non-Abcam antibody was used: anti-rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution: 1/20000

Detection method: west pico
Exposure: 30 second(s)
Bands: Specific: 4 kDa

Additional data
Additional Notes: Band was detected at 4kDa but should appear at 1.7kDa. Probably due to uncertain running of a very low MW product. Only 1 band should be present in the sample and only one band was detected.
Abcam response: Thank you for submitting your Abreview. While you have been able to detect the 1.4 kDa band, we would not typically recommend this peptide for use as a WB positive control because it is very difficult to successfully run and transfer peptides of this size. The peptide is commonly used for blocking purposes.

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Submitted Aug 01 2012

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