Anti-TATA binding protein TBP antibody [58C9] - Loading Control (ab61411)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Mouse (1:4000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Mouse (1:4000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

TCA extracts were prepared from 10 OD600 units of log phase and separated quiescent and non-quiescent cells isolated from at least 2–3 independent experiments [51]. A sonication step of 4 × 10 s was added after glass-bead lysis and collection in 500 μL 5% TCA. 10 – 50 μg of protein were separated on 10 or 15% polyacrylamide gels, blotted to an Immobilon-P membrane, and incubated with the antibodies described in the following section.

Antibodies
Western blots were incubated with the following antibodies: anti-FLAG (Sigma F3165: 1/15,000); anti-beta actin (Abcam ab8224: 1/3000–10,000); anti-Myc (Millipore 05–419, 9E10 monoclonal: 1/2000); anti-H2A (Active Motif 39235: 1/3000-10,000); anti-H2A.Z (Active Motif 39647: 1/1000-2000); anti-H2B (Active Motif 39237: 1/2500-10,000); anti-H3 (Active Motif 39163: 1/5000-7500; anti-H4 (Abcam ab7311: 1/1000); anti-H3K9/K14Ac (Millipore 06–599: 1/5000); anti-H3K9ac (Millipore 07–352: 1/5000); anti-H3K14ac (Millipore 07–353: 1/5000); anti-H4tetraAc (Millipore 06–886: 1/2500-6000); anti-H4K16ac (Active Motif 39929: 1/2000); anti-H3K56ac (Millipore 07-677-I: 1/1000-2000); anti-H3K4me1 (Millipore 07–436: 1/2000); anti-H3K4me2 (Active Motif 39913: 1/5000 and Abcam ab7766: 1/1000); anti-H3K4me3 (Active Motif 39159: 1/3000-5000); anti-H3K36me2 (Active Motif 39255: 1/5000); anti-H3K36me3 (Abcam ab9050: 1/1000-3000); anti-H3K79me1 (Abcam ab2886: 1/4000 and Active Motif 39145: 1/2000); anti-H3K79me2 (Millipore 04–835: 1/2000-3000); anti-H3K79me3 (Abcam ab2621: 1/7000-10,000); H14, anti-CTD-Ser5P (Covance MMS134R: 1/1000 and Active Motif 39750: 1/1000); H5, anti-CTD-Ser2P (Covance MMS-129R: 1/500-2000); anti-Gcn5 (Santa Cruz sc-9078: 1/500); anti-Rpd3 (Santa Cruz sc-6655: 1/500); anti-Hda1 (Santa Cruz sc-9077: 1/500); anti-Sir2 (Santa Cruz sc-6666: 1/500); anti-TBP (Abcam ab61411: 1/4000); and anti-Srb4 (Abcam ab63812: 1/5000).

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Manufacturer protocol

Download the product protocol from Abcam for Anti-TATA binding protein TBP antibody [58C9] - Loading Control (ab61411) below.

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