Figure 4. Proteomic survey of mouse tissues with azo-resin E.

Tissue extracts were prepared from the indicated tissues and applied to a fixed volume of resin E (100μl) in the absence (A) and presence (B) of 5 (100 μM). Following washes at physiological ionic strength the bound proteins were eluted with 30 mM dithionite and characterized by SDS-PAGE (4–15% acrylamide), silver staining and mass spectrometry. Key: 1–22. full length Hsp90α/β or N terminal fragments of Hsp90α/β; 23. epoxide hydroxylase; 24. glutaryl-CoA dehydrogenase; 25. and 26. Hsp90α/β; 27. Hsp70; 28. epoxide hydroxylase; 29. delta(3,5)-delta(2,4)-dienoyl-CoA isomerase; 30. 3,2-trans-enoyl-CoA isomerase/NADPH-dependent retinol reductase; 31 and 32. Hsp90α/β; 33. Hsp70; 34. Hsp90α/β; 35. glycogen debranching enzyme; 36 and 37. Hsp90α/β; 38. epoxide hydroxylase; 39. 3-ketoacyl-CoA thiolase/3-ketoacyl-CoA thiolase; 40–42. Hsp90α/β; 43. Hsp70 44. epoxide hydroxylase/liver carboxylesterase 31; 45–47 Hsp90α/β; 48. epoxide hydroxylase; 49–51. Hsp90α/β; 52. Albumin; 53. Hsp90α/β; 55. Albumin; 56. CAZ-associated structural protein 1/Ankrd11 protein; 56. Hsp90α/β; 57. Albumin; 58. epoxide hydroxylase; 59. Glutaryl-CoA dehydrogenase; 60. Hsp90α/β. Combined consisted of a mixture of striated muscle, liver, testis and adipose tissue. The lane w/o dithionite was eluted with SDS. This gel is a representative example of using resin E against tissue extracts. (A larger view is shown in the supplemental material, Supplementary Fig. S5)