Protein content varies considerably across distinct biological input material (for example different cell lines, strains, tissues, tissue regions, biological fluids and the sample storage conditions the samples are subjected to (e.g. different storage temperatures, storage time, repeated freeze-thaw cycles,fresh frozen vs. FFPE, formaldehyde treatment). We advise to quantify protein concentration of your sample after the lysis step (see "Shall I adjust the buffer volumes depending on the protein starting amounts?"). A short overview of raw material amounts is given in the table below.
We have a database with processing recommendations for >100 species and sample matrices. Please
contact us for further details on your specific sample of interest
Material
Starting amount
Protein amount
Mammalian Cell Line (e.g. HeLa)
6E5 cells
100 µg
Yeast (S.cerevisae)
OD600 = 0.6
100 µg
Bacteria (E.coli)
OD600 = 0.5
100 µg
Immunoprecipitation
1 mL slurry
10 - 400 µg
Blood / Serum / Plasma (H.sapiens)
2 µL
100 µg
Mammalian Tissue
1 mm3
100 µg
Plant Tissue (A.thaliana): shoot/root wet weight
50 mg / 100 mg
100 mg / 100 mg