Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-It is highly critical that the chromatin is of appropriate size and concentration.	-It is important to keep the tissue cold to avoid protein degradation. -Use fresh formaldehyde that is not past the manufacturer's expiration date.	-Once in solution, store 1M DTT at -20°C.

Upstream tips
-It is highly critical that the chromatin is of appropriate size and concentration.
Protocol tips
-It is important to keep the tissue cold to avoid protein degradation. -Use fresh formaldehyde that is not past the manufacturer's expiration date.
Downstream tips
-Once in solution, store 1M DTT at -20°C.

Publication protocol

ChIP was performed using the Magnify Chromatin Immunoprecipitation System (Invitrogen), according to the manufacturer’s instructions. Cells transfected with PC4-Flag or Flag empty vector, treated with or without TSA, were collected and washed with 1x PBS, and then fixed with 1% formaldehyde for 10 min. Chromatin was then prepared by sonication shearing, optimized according to the manufacturer’s instructions. ChIP was performed on sheared chromatin from approximately 2 × 105 cells, using IgG antibody as a negative control or antibodies recognizing Flag, TFIIB, Pol II, H3K9-Ac, or H3.3. Chromatin purified from an equivalent number of cells not subjected to the immunoprecipitation step was used as the input control. Real-time PCR was performed on DNA isolated from each of the ChIP reactions using primer pairs specific for LHR 5′-ACTGGGCACTGTCGCAGGTC-3′ (sense) and 5′-CATGGCCGGCGAACTGGGCT-3′ (anti-sense) [6]. The amplified region of the 189 bp LHR promoter contains the relevant proximal Sp1 response element.

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Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing ChIP Anti-bodies H3K9-Ac using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 from Cell Signaling Technology.

Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 below.

Download manufacturer protocol

Videos

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