Incubation:Article Title: Structure of the Large Extracellular Loop of FtsX and Its Interaction with the Essential Peptidoglycan Hydrolase PcsB in Streptococcus pneumoniae
Article Snippet: .. Samples were diluted with 2X 246 Laemlli SDS loading buffer (Bio-Rad) and incubated at 95°C for 10 min. 12.5 μg of total 247 protein was loaded per sample onto a 4-15% precast gradient SDS-PAGE gel (Bio-Rad, 248 4561084) and subjected to electrophoresis for 1 hour at 150 V. Proteins were 249 transferred to a nitrocellulose membrane and probed with polyclonal rabbit α-FtsX 250 12 (1:500, see experimental procedures for production) primary antibody and ECL anti-251 rabbit IgG horseradish peroxidase linked whole antibody (1:10,000, GE Healthcare, 252 NA93AV) secondary antibody in TBST (Tris-Buffered Saline, 0.02% Tween). .. Antibody 253 signal was detected by incubating membranes with Amersham ECL Western Blotting 254 Detection Reagent (GE Healthcare, RPN2106) and detecting chemiluminescent signals 255 from protein bands using an IVIS imaging system with a 1 min exposure as described in 256 previously (9).
SDS Page:Article Title: Structure of the Large Extracellular Loop of FtsX and Its Interaction with the Essential Peptidoglycan Hydrolase PcsB in Streptococcus pneumoniae
Article Snippet: .. Samples were diluted with 2X 246 Laemlli SDS loading buffer (Bio-Rad) and incubated at 95°C for 10 min. 12.5 μg of total 247 protein was loaded per sample onto a 4-15% precast gradient SDS-PAGE gel (Bio-Rad, 248 4561084) and subjected to electrophoresis for 1 hour at 150 V. Proteins were 249 transferred to a nitrocellulose membrane and probed with polyclonal rabbit α-FtsX 250 12 (1:500, see experimental procedures for production) primary antibody and ECL anti-251 rabbit IgG horseradish peroxidase linked whole antibody (1:10,000, GE Healthcare, 252 NA93AV) secondary antibody in TBST (Tris-Buffered Saline, 0.02% Tween). .. Antibody 253 signal was detected by incubating membranes with Amersham ECL Western Blotting 254 Detection Reagent (GE Healthcare, RPN2106) and detecting chemiluminescent signals 255 from protein bands using an IVIS imaging system with a 1 min exposure as described in 256 previously (9).
Electrophoresis:Article Title: Structure of the Large Extracellular Loop of FtsX and Its Interaction with the Essential Peptidoglycan Hydrolase PcsB in Streptococcus pneumoniae
Article Snippet: .. Samples were diluted with 2X 246 Laemlli SDS loading buffer (Bio-Rad) and incubated at 95°C for 10 min. 12.5 μg of total 247 protein was loaded per sample onto a 4-15% precast gradient SDS-PAGE gel (Bio-Rad, 248 4561084) and subjected to electrophoresis for 1 hour at 150 V. Proteins were 249 transferred to a nitrocellulose membrane and probed with polyclonal rabbit α-FtsX 250 12 (1:500, see experimental procedures for production) primary antibody and ECL anti-251 rabbit IgG horseradish peroxidase linked whole antibody (1:10,000, GE Healthcare, 252 NA93AV) secondary antibody in TBST (Tris-Buffered Saline, 0.02% Tween). .. Antibody 253 signal was detected by incubating membranes with Amersham ECL Western Blotting 254 Detection Reagent (GE Healthcare, RPN2106) and detecting chemiluminescent signals 255 from protein bands using an IVIS imaging system with a 1 min exposure as described in 256 previously (9).
Membrane:Article Title: Structure of the Large Extracellular Loop of FtsX and Its Interaction with the Essential Peptidoglycan Hydrolase PcsB in Streptococcus pneumoniae
Article Snippet: .. Samples were diluted with 2X 246 Laemlli SDS loading buffer (Bio-Rad) and incubated at 95°C for 10 min. 12.5 μg of total 247 protein was loaded per sample onto a 4-15% precast gradient SDS-PAGE gel (Bio-Rad, 248 4561084) and subjected to electrophoresis for 1 hour at 150 V. Proteins were 249 transferred to a nitrocellulose membrane and probed with polyclonal rabbit α-FtsX 250 12 (1:500, see experimental procedures for production) primary antibody and ECL anti-251 rabbit IgG horseradish peroxidase linked whole antibody (1:10,000, GE Healthcare, 252 NA93AV) secondary antibody in TBST (Tris-Buffered Saline, 0.02% Tween). .. Antibody 253 signal was detected by incubating membranes with Amersham ECL Western Blotting 254 Detection Reagent (GE Healthcare, RPN2106) and detecting chemiluminescent signals 255 from protein bands using an IVIS imaging system with a 1 min exposure as described in 256 previously (9).
Saline:Article Title: Structure of the Large Extracellular Loop of FtsX and Its Interaction with the Essential Peptidoglycan Hydrolase PcsB in Streptococcus pneumoniae
Article Snippet: .. Samples were diluted with 2X 246 Laemlli SDS loading buffer (Bio-Rad) and incubated at 95°C for 10 min. 12.5 μg of total 247 protein was loaded per sample onto a 4-15% precast gradient SDS-PAGE gel (Bio-Rad, 248 4561084) and subjected to electrophoresis for 1 hour at 150 V. Proteins were 249 transferred to a nitrocellulose membrane and probed with polyclonal rabbit α-FtsX 250 12 (1:500, see experimental procedures for production) primary antibody and ECL anti-251 rabbit IgG horseradish peroxidase linked whole antibody (1:10,000, GE Healthcare, 252 NA93AV) secondary antibody in TBST (Tris-Buffered Saline, 0.02% Tween). .. Antibody 253 signal was detected by incubating membranes with Amersham ECL Western Blotting 254 Detection Reagent (GE Healthcare, RPN2106) and detecting chemiluminescent signals 255 from protein bands using an IVIS imaging system with a 1 min exposure as described in 256 previously (9).
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