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edits
Changes
added protocol for tissue extraction
==Materials==
EpiQuik Nuclear Extraction Kit [https://www.epigentek.com/catalog/epiquik-nuclear-extraction-kit-p-149.html Epigentek OP-0002-1] 100 Extractions $184.00
Epigenase HDAC Activity/Inhibition Direct Assay Kit (Colorimetric) [https://www.epigentek.com/catalog/epigenase-hdac-activityinhibition-direct-assay-kit-colorimetric-p-2867.html Epigentek P-4034-96] 96 assays $415.00
==Tissue Nuclear Extraction==
===For Tissue samples===
1. Weigh tissue and cut it into small pieces.
2. Add to 2ml homogenizer tube with NE1 buffer (5000 ul/mg of tissue)
3. Homogenize (50-60 strokes).
4. Incubate on ice for 15mins
5. Centrifuge for 10mins at 12000 rpm at 4C
6. Remove the supernatant
===Nuclear Extract Preparation===
1. Add 2 volumes (based on pellet size) of NE2 containing DTT and PIC to nuclear pellet (2500 ul / mg of initial tissue)
2. Incubate the extract on ice for 15mins with vortex (5secs) every 3 mins. (The tissue extract can be further sonicated for 3 x 10 seconds to increase nuclear protein extraction).
[[Category: CTA Experiments]]
3. Centrifuge the suspension for 10mins at 14000 rpm at 4C and transfer the supernatant into a new micro-centrifuge vial.
4. Use immediately or aliquot and freeze supernatant at -80C until further use.
Be sure to save aliquot for protein concentration.
5. Measure the protein concentration of the nuclear extract
[[Category: Epigenetics]]