Tissue Protein Extraction Buffer

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Applikation
Protein Extraction (PEx)
Optionen
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Produktmerkmale Extracts approximately 90% of proteins from tissues, including transmembrane proteins.
Simple and rapid protocol for tissue protein extraction. Use mini-homogenizer-pestle, microcentrifuge tubes and table top microcentrifuge in the protocol.
Maintains protein-protein complexes for downstream immunoprecipitation and pull-down experiments. ELISA compatible. Non-denaturing.
Can be applied directly to extract tissue proteins for Western blotting.
Compatible with Bradford assay.
Absence of amines in the buffer allows for NHS-Ester conjugation and biotinylation.
Applikationshinweise Sufficient for 20 mL minced tissue.
Testdurchführung

Protocol (Keep solutions on ice):
Pulverize approximately 90 µL of tissue. Place tissue in a 1.5 mL round bottom microcentrifuge tube.
Add general phosphatase and protease inhibitor cocktails to 500 µL of ice-cold Tissue Protein Extraction Buffer.
Add 500 µL Tissue Protein Extraction buffer with inhibitors to pulverized tissue.
Homogenize tissue with a mini pestle-homogenizer using 15 strokes, 3 seconds/stroke on ice.
Centrifuge 12000g for 15 min at 4oC.
Remove supernatant (without lipid layer) and transfer into another 1.5 mL tube.
Centrifuge again at 12000g for 15 min at 4oC.
Transfer supernatant to another tube. The supernatant fraction contains the extracted proteins.
The Bradford assay can be used to quantitate extracted protein concentration.

Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Lagerung -20 °C
Produkt verwendet in: Chiu, Hu, Yang, Hsin, Ko, Tsai, Sheu: "Immunomodulatory Protein from Ganoderma microsporum Induces Pro-Death Autophagy through Akt-mTOR-p70S6K Pathway Inhibition in Multidrug Resistant Lung Cancer Cells." in: PLoS ONE, Vol. 10, Issue 5, pp. e0125774, 2015 (PubMed).