Owner manual

cell). Expected protein recovery using this kit is approximately
20% (based on studies using 100 mg rat brain tissue,
mouse liver tissue, and Jurkat cells), and the extraction
process requires approximately 1 hr.
Each MicroRotofor run using 2.5 mg total protein yields ten
150–250 µl fractions, and the protein distribution among
the fractions will vary depending on the protein sample. For
example, using rat brain samples and ampholytes spanning
the pH range 3–10, fractions 3–7 typically contain the most
protein. Fractions are cleaned up using the ReadyPrep
™
2-D cleanup kit, then are ready for downstream applications
(Harbers et al. 2005). As a starting point, 50 µl of a fraction
is generally enough to run triplicate 2-D gels using 11 cm
IPG strips and stained with Flamingo
™
fluorescent stain
(catalog #161-0490) for visualization.
Certificates of analysis and MSDS forms are available upon
request.
Items Supplied With Kit
Protein solubilization buffer (PSB) (contains urea, 25 g
thiourea, NDSB 201, and Tris)
PSB diluent (contains CHAPS, Tris) 30 ml
ReadyPrep mini grinders 2 packets
of 10
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