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. Author manuscript; available in PMC: 2013 May 1.
Published in final edited form as: Methods Mol Biol. 2012;917:369–390. doi: 10.1007/978-1-61779-992-1_21

Table 2.

Examples of lysis buffers used for immunoaffinity purification of protein complexes

Bait
(GFP-tagged)		 Description
and localization		 Species
and sample type		 Optimized lysis buffer References
Apl1 beta-adaptin of AP-2 complex; cellular membrane S. cerevisiae cells 20 mM K-HEPES, pH 7.4, 110 mM KOAc, 2 mM MgCl2, 0.1% Tween, 0.4% Triton, 200 mM NaCl, 1/100 (v/v) protease inhibitor mixture (20 mg/mL PMSF + 0.4 mg/mL pepstatin A), and 1/200 (v/v) protease inhibitor cocktail (2)
Nup37 Member of nuclear pore complex; subunit of Nup107–160 subcomplex Homo sapiens cells 20 mM K-HEPES, pH 7.4, 110 mM KOAc, 2 mM MgCl2, 0.1% TWeen, 0.5% Triton, 200 mM NaCl, 1/100 (v/v) protease inhibitor mixture (20 mg/mL PMSF + 0.4 mg/mL pepstatin A), and 1/200 (v/v) protease inhibitor cocktail (2)
HDAC5 Histone deacetylase 5; nucleus and cytoplasm Homo sapiens cells 20 mM HEPES-KOH, pH 7.4, 0.1 M potassium acetate, 2 mM MgCl2, 0.l% Tween-20, 1 mM ZnCl2, 1 mM CaCl2, 0.5% Triton X-100, 250 mM NaCl, 4 mg/ml DNase, 1/100 (v/v) protease, and phosphatase inhibitor cocktails (7)
PAP I poly (A) polymerase I; cytoplasm and inner membrane Escherichia coli cells 20 mM HEPES, pH 7.4, 0.11 M KOAc, 2 mM MgCl2, 0.1% Tween-20 (v/v), 1 mM ZnCl2 1 mM CaCl2, 1% Triton X-100, 0.5% Deoxycholate, 150 mM NaCl, 1/100 protease inhibitor cocktail, 1/200 phenylmethylsulphonyl fluoride (6)
H3 Histone 3 isoforms; nucleus Mus musculus ES cells 20 mM K-HFPES, pH 7.4, 110 mM K-acetate, 0.1% Tween 20, 0.5% Triton, 300 mM NaCl, and 1/100 (v/v) protease inhibitor cocktail (5)
nsP3 Sindbis nonstructural protein 3; cytoplasm Sindbis-infected Rattus Norvegicus cells 20 mM K-HEPES, pH 7.4, 110 mM KOAc, 2 mm MgCl2, 0.1% Tween 20, 1% Triton, 0.5% deoxycholate, 500 mM NaCl, 25 units/mL DNase, 1/100 (v/v) protease inhibitor mixture (3)
PSD (via VGluRd2) postsynaptic: densities; cerebellar excitatory synapses Mus musculus tissue 10 mM HEPES, pH 7.4, 2 mM CaCl2, 132 mM NaCl, 3 mM KCl, 2 mM MgSO4, 1.2 mM NaH2PO4, 0.5% Triton X-100, 1/100 (v/v/) protease inhibitor cocktail (8)