Table 3.

Classification of mtDod-PAS-Pep derived ABs. ABs were classified as followed: “Class 1” ABs recognize the proteins of interest provided as recombinantly purified protein (produced in E. coli). “Class 2” ABs do not recognize recombinant protein of interest, but protein of expected apparent molecular weight in HEK293T human cell lysates. “Class 3” ABs only recognize mtDod-PAS constructs (like the respective mtDod-PAS-Pep or mtDod-PAS-Met). Western blots of ABs rated as “class 1” or “class 2” are shown in Fig. 8. The AB derived from mtDod-PAS-Pep6 and mtDod-PAS-Pep8 only recognized mtDod-PAS-Pep constructs and were considered “class 3” (Supplementary Fig. S11).

ABs derived from mtDod construct	Binding region of the ABs	AB class
mtDod-PAS-Pep1	HSP-70 C-terminus	Class 1
mtDod-PAS-Pep2	HSP-90 C-terminus	Class 1
mtDod-PAS-Pep3	HSP-110 C-terminus	Class 1
mtDod-PAS-Pep4	HSP-A4 C-terminus	Class 1
mtDod-PAS-Pep5	H3 C-terminus	Class 2*
mtDod-PAS-Pep6	H4 C-terminus	Class 3
mtDod-PAS-Pep8	HB-EGF** Pos. 20–34	Class 3
mtDod-PAS-Pep9	CHIP N-terminus	Class 1
mtDod-PAS-Pep10	CHIP Broken helix Pos. 151–161	Class 2***
mtDod-PAS-Pep11	CHIP Tip of helix Pos. 183–192	Class 1

*mtDod-PAS-Pep5 derived ABs showed only a very weak signal for 1 µg and 500 ng of recombinant protein with no intensity difference (see Supplementary Fig. S11). Thus, the AB preparation was regarded as “class 2”.

**Proheparin-binding EGF-like growth factor of Chlorocebus aethiops (green monkey).

***mtDod-PAS-Pep10 derived ABs didn’t recognize purified CHIP but seem to recognize a protein in CHIP-overexpressing cells; no detection range was determined.