Membrane:Article Title: Polymeric assembly of endogenous Tuberous Sclerosis Protein Complex
Article Snippet: .. The membranes were washed again in TBST buffer for 5 minutes three times before incubating overnight for 17 hours in their respective primary antibody solutions: the first membrane was incubated in 10 mL rabbit anti-TSC1 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TSC1 antibody (Cell Signaling Technology, Clone #D43E2, Lot #3)], the second membrane was incubated in 10 mL rabbit anti-TSC2 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v rabbit anti-TSC2 antibody (Cell Signaling Technology, Clone #D93F12, Lot #6)] and the last membrane was incubated in 10 mL rabbit anti-TBC1D7 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TBC1D7 antibody (Cell Signaling Technology, Clone #D8K1Y, Lot #3)]. .. The membranes were then washed in TBST buffer for 5 minutes three times then incubated for 1 hour at room temperature in their respective secondary antibody solutions: the membranes blotted with rabbit anti-TSC1 and rabbit anti-TSC2 primary antibodies were incubated in 10 mL goat anti-rabbit secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit secondary antibody (Cell Signaling Technology, Product #7076S, Lot #33)] and the membrane blotted with rabbit anti-TBC1D7 primary antibody was incubated in 10 mL goat anti-rabbit F c secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit F c secondary antibody (Abcam, Lot #GR3347004-5)].
Article Title: Polymeric assembly of endogenous Tuberous Sclerosis Protein Complex
Article Snippet: .. The membranes were washed again in TBST buffer for 5 minutes three times before incubating overnight for 17 hours in their respective primary antibody solutions: the first membrane was incubated in 10 mL rabbit anti-TSC1 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TSC1 antibody (Cell Signaling Technology, Clone #D43E2, Lot #3)], the second membrane was incubated in 10 mL mouse anti-TSC1 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v mouse anti-TSC2 antibody (Invitrogen, Ref #37-0400, Lot #UD288308)], the third membrane was incubated in 10 mL rabbit anti-TSC2 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v rabbit anti-TSC2 antibody (Cell Signaling Technology, Clone #D93F12, Lot #6)] and the fourth membrane was incubated in 10 mL rabbit anti-TBC1D7 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TBC1D7 antibody (Cell Signaling Technology, Clone #D8K1Y, Lot #3)]. .. The membranes were then washed in TBST buffer for 5 minutes three times then incubated for 1 hour at room temperature in their respective secondary antibody solutions: the membranes blotted with rabbit anti-TSC1 and rabbit anti-TSC2 primary antibodies were incubated in 10 mL goat anti-rabbit secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit secondary antibody (Cell Signaling Technology, Product #7074P2, Lot #28)], the membrane blotted with mouse anti-TSC1 primary antibody was incubated in 10 mL goat anti-mouse secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-mouse secondary antibody (Cell Signaling Technology, Product #7076S, Lot #33)] and the membrane blotted with anti-TBC1D7 primary antibody was incubated in 10 mL goat anti-rabbit F c secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit F c secondary antibody (Abcam, Lot #GR3347004-5)].
Incubation:Article Title: Polymeric assembly of endogenous Tuberous Sclerosis Protein Complex
Article Snippet: .. The membranes were washed again in TBST buffer for 5 minutes three times before incubating overnight for 17 hours in their respective primary antibody solutions: the first membrane was incubated in 10 mL rabbit anti-TSC1 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TSC1 antibody (Cell Signaling Technology, Clone #D43E2, Lot #3)], the second membrane was incubated in 10 mL rabbit anti-TSC2 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v rabbit anti-TSC2 antibody (Cell Signaling Technology, Clone #D93F12, Lot #6)] and the last membrane was incubated in 10 mL rabbit anti-TBC1D7 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TBC1D7 antibody (Cell Signaling Technology, Clone #D8K1Y, Lot #3)]. .. The membranes were then washed in TBST buffer for 5 minutes three times then incubated for 1 hour at room temperature in their respective secondary antibody solutions: the membranes blotted with rabbit anti-TSC1 and rabbit anti-TSC2 primary antibodies were incubated in 10 mL goat anti-rabbit secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit secondary antibody (Cell Signaling Technology, Product #7076S, Lot #33)] and the membrane blotted with rabbit anti-TBC1D7 primary antibody was incubated in 10 mL goat anti-rabbit F c secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit F c secondary antibody (Abcam, Lot #GR3347004-5)].
Article Title: Polymeric assembly of endogenous Tuberous Sclerosis Protein Complex
Article Snippet: .. The membranes were washed again in TBST buffer for 5 minutes three times before incubating overnight for 17 hours in their respective primary antibody solutions: the first membrane was incubated in 10 mL rabbit anti-TSC1 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TSC1 antibody (Cell Signaling Technology, Clone #D43E2, Lot #3)], the second membrane was incubated in 10 mL mouse anti-TSC1 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v mouse anti-TSC2 antibody (Invitrogen, Ref #37-0400, Lot #UD288308)], the third membrane was incubated in 10 mL rabbit anti-TSC2 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v rabbit anti-TSC2 antibody (Cell Signaling Technology, Clone #D93F12, Lot #6)] and the fourth membrane was incubated in 10 mL rabbit anti-TBC1D7 primary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:1000 v/v rabbit anti-TBC1D7 antibody (Cell Signaling Technology, Clone #D8K1Y, Lot #3)]. .. The membranes were then washed in TBST buffer for 5 minutes three times then incubated for 1 hour at room temperature in their respective secondary antibody solutions: the membranes blotted with rabbit anti-TSC1 and rabbit anti-TSC2 primary antibodies were incubated in 10 mL goat anti-rabbit secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit secondary antibody (Cell Signaling Technology, Product #7074P2, Lot #28)], the membrane blotted with mouse anti-TSC1 primary antibody was incubated in 10 mL goat anti-mouse secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-mouse secondary antibody (Cell Signaling Technology, Product #7076S, Lot #33)] and the membrane blotted with anti-TBC1D7 primary antibody was incubated in 10 mL goat anti-rabbit F c secondary antibody solution [50 mM Tris, 100 mM NaCl, 1% v/v Tween 20, 5% w/v skim milk powder, 1:5000 v/v goat anti-rabbit F c secondary antibody (Abcam, Lot #GR3347004-5)].
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