View source for HDAC Activity Sex Differences


==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).

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]]