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Rat Liver Preparation
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Materials
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Ordering information
Related product information
ELISA Kits
Quant-iT Protein Kits for Microplates
Qubit Protein Quantitation
Materials
Aluminum Foil
Liquid Nitrogen
Dry Ice
5 mL Culture Tubes
Phosphate Buffered Saline
(Cat. No. 20012050)
Protease Inhibitor Cocktail
(Cat. No. 78429)
BCA Protein Assay or Quant-iT Protein Quantitation Kit
(Cat. No. Q33210)
Protocol
Remove liver aseptically.
Flash-freeze liver in foil packets using liquid nitrogen. Store at –80°C.
Crush liver sample on dry ice and store in a prechilled 5 mL culture tube.
Add about 100 μg tissue (enough to almost cover round portion of a 5 mL culture tube) to a new chilled tube.
Add 1 mL PBS with 10 μL protease inhibitors. Chill this solution using wet ice.
Homogenize at low speed for ~20 seconds. Make sure to keep cool and keep samples on wet ice.
Transfer samples into 1.7 mL microcentrifuge tubes.
Centrifuge at 14,000 x g at 4°C for 15 minutes.
Remove and aliquot supernatant. We recommend making several 50 μL aliquots.
Before running ELISA, dilute protein 1:100 and run a
BCA total protein assay
or Quant-iT Protein Quantitation Kit assay (Cat. No. Q33210).
Dilute the samples to equal concentrations and then run ELISA as described under
ELISA Protocol
.
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BioSource C-070276 1107 1-Jan-2007
Ordering information
Q33210