Comparison of Flow Cytometry and Western Blotting To Measure Hsp70
作者： Liusheng He , Michael H. Fox
DOI: 10.1002/(SICI)1097-0320(19961101)25:3<280::AID-CYTO9>3.0.CO;2-J
关键词: Chinese hamster ovary cell 、 Hsp70 、 Blot 、 Antibody 、 Cell cycle 、 Molecular biology 、 Flow cytometry 、 Chemistry 、 Propidium iodide 、 Protein tyrosine phosphatase
摘要: The levels of constitutive and inducible forms heat shock protein 70 (hsp73 hsp72, respectively) through the cell cycle were measured in CHO cells by flow cytometry Western blotting at various times after heating. Cells labeled with antibody C92 (hsp72) or N27 (hsp73) propidium iodide prior to analysis cytometry. heated for 15 min 45 degrees C, then analyzed from 3 36 h later. There was about a tenfold increase hsp72 early S phase beginning within 6 heating these gradually cycled though so most them had divided. When exposed 10 microM sodium vanadate, an inhibitor tyrosine phosphatase, 24 heating, induction almost completely inhibited. Heated did not express hsp73 cell-cycle-dependent manner. Hsp73 increased uniformly all vanadate affect expression. Quantitative comparisons relative excellent agreement. Control Hoechst 33342 sorted G1, S, G2/M phases processed verify dependent as Again there agreement between results.
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