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added protocol for tissue extraction
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+[[Category: Taste Aversion]]+
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==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]]