Measurement of Autophagy in Cells and Tissues - Protein Misfolding and Cellular Stress in Disease and Aging - page 194

Table 2
Antibodies suitable for recognizing endogenous autophagy-related proteins
by Western blotting

Name Information/company/conditions for western blotting

Anti-Atg3 (1) Rabbit polyclonal antibody, Sigma-Aldrich, A3231;

blocking, 5% BSA -TBS, incubate at room tem-
perature for 30 min; primary antibody reaction
(dilution, 1:2,000), incubate at room temperature
for 90-120 min

(2) Mouse monoclonal IgG2b kappa, Medical &
Biological Laboratories, Tokyo, Japan, #M133-3;
blocking, 10% skim milk-TBS, incubate at room
temperature for 30 min; primary antibody reaction
(dilution, 1:2,000), incubate at 4 ° C for 15 h

Anti-Atg4B (1) Rabbit polyclonal antibody, Sigma-Aldrich, A2981;

blocking, 5% BSA -TBS, incubate at room tem-
perature for 30 min; primary antibody reaction
(dilution, 1:2,000), incubate at room temperature
for 90-120 min

Anti-Atg5 (1) Rabbit polyclonal antibody, Sigma-Aldrich, A0731;

blocking, 5% BSA -TBS, incubate at room tem-
perature for 30 min; primary antibody reaction
(dilution, 1:2,000), incubate at room temperature
for 90-120 min

(2) Rabbit polyclonal antibody specific for human
Atg5 , Cell Signaling Technology, Danvers, MA,
#2630; blocking, 1% BSA (or 10% skim milk)-TBS,
incubate at room temperature for 30 min; primary
antibody reaction (dilution, 1:2,000), incubate at
room temperature for 90-120 min

(3) Chicken polyclonal antibody, Abcam, Cambridge,
MA, ab19130; blocking, 1% BSA (or 10% skim
milk)-TBS, incubate at room temperature for
30 min; primary antibody reaction (dilution,
1:2,000), incubate at 4 ° C for 15 h

Anti-Atg7 (1) Rabbit polyclonal antibody, Sigma-Aldrich, A2856;

blocking, 5% BSA -TBS, incubate at room
temperature for 30 min; primary antibody reaction
(dilution, 1:2,000), incubate at room temperature
for 90-120 min

(2) Rabbit polyclonal antibody, Cell Signaling
Technology, #2631; blocking, 1% BSA (or 10%
skim milk)-TBS, incubate at room temperature for
30 min; primary antibody reaction (dilution,
1:2,000), incubate at room temperature for
90-120 min

(continued)