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, Fetal bovine serum (FBS) (Lonza)

, DMEM w/FBS: DMEM supplemented with 10% FBS

, Cell culture grade trypsin solution (Lonza)

, Ethylenediaminetetraacetic acid (EDTA): 1% (Sigma)

, Sterile washing buffer: HBSS (Lonza)

, Transfection reagent: DreamFectTM Gold ® (Oz biosciences)

D. and /. Fbs-10%,

, Sterile washing buffer: HBSS (Lonza)

-. Opti and . Medium,

. Human, HA hHSP27 or hHSP70

, Sterile washing buffer: HBSS (Lonza)

D. and /. Fbs-10%,

, Phosphate-buffered saline (PBS) 1× (Lonza)

, Lysis buffer: 50 mM HEPES, 0.25 M EDTA, 4 M NaCl, 0.1% NP40

, Antiprotease: Prepare in advance in sterile water. Store at ?20°C

, Lowry protein assay

, Sonicator. 1. 96-well plate (flat bottom)

, BSA: Prepare in advance at 100 g/L, store at ?20°C

, DTT: Prepare in advance at 1 M, store at ?20°C

, Sodium phosphate buffer at 50 mM

, Hsp27 recombinant protein: Store separate aliquots at ?80°C (see Note 2)

, For protease inhibition, add 300 mL of antiprotease to 10 mL lysis buffer. Keep the buffer on ice (see Note 11)

, Add 300 mL of lysis buffer (see Note 12)

, Transfer in microtube and vortex for 10 s. Keep on ice during 20 min for cell lysis

, Sonicate for 10 s, power 10%

, Adjust protein concentration of all samples at 2 mg/mL with sodium phosphate buffer or Tris-HCl buffer

, Samples can be stored at ?80°C supplemented with 3% of glycerol before aggregation test (see Note 14)

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