ChIP Anti-bodies H3K27me3

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product is observed in the experimental sample, add more DNA to the PCR reaction or increase the number of amplification cycles. Furthermore, if you have any problem with antibodies, make sure to use the ChIP-validated antibody.

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Found 3 matching solutions for this experiment

Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733

Cell Signaling Technology

Upstream tips	Protocol tips	Downstream tips
-It is highly critical that the chromatin is of appropriate size and concentration.	-For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. -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
-For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. -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.

Manufacturer protocol Publication protocol 1 Relevant paper

Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729)

Abcam

Upstream tips	Protocol tips	Downstream tips
-Add protease inhibitors to all lysis solutions before use.	-Use 2 µg for 25 µg of chromatin. -Keep cells on ice between the rounds of homogenisations. - Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line.

Upstream tips
-Add protease inhibitors to all lysis solutions before use.
Protocol tips
-Use 2 µg for 25 µg of chromatin. -Keep cells on ice between the rounds of homogenisations. - Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line.

Manufacturer protocol Publication protocol 1 Relevant paper

Anti-trimethyl-Histone H3 (Lys27) Antibody

Millipore

Upstream tips	Protocol tips	Downstream tips
	The ChIP assay was performed using the imprint chromatin immunoprecipitation kit (CHP1, Sigma-Aldrich) following the manufacturer's instructions. The sonicated DNA was incubated with the specific antibody.

Protocol tips
The ChIP assay was performed using the imprint chromatin immunoprecipitation kit (CHP1, Sigma-Aldrich) following the manufacturer's instructions. The sonicated DNA was incubated with the specific antibody.

Manufacturer protocol Publication protocol 1 Relevant paper

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