On June 3, 2015Christian Kroun Damgaard, Aarhus University wrote:
4.3
Western using lysates from HeLa, NHDF, HEK293
1:2000 for western (standard 5% milk, TBS-T buffer).
Secondary antibodies dilution: 1:5.000 of goat anti-rabbit/HRP antibodies in 5%milk/TBS-T (DAKO #P0448) (incubation time: 1hr at RT)
Chemiluminescent substrate: SuperSignal West Femto (Thermo Scientific #34096)
Clean and robust band at approximately 70 kDa detected.
Immunofluorescence using HeLa, NHDF, HEK293 cells fixed in 4% PFA (PBS), Permeabilization in 0.5% Triton/PBS/1% horse serum). Primary: 1:1000 dilution in PBS/1% horse serum. Secondary: 1:1000 dilution PBS/1% horse serum. Donkey anti rabbit – Alexa 488 or Alexa 594 conjugated. Bright cytoplasmic staining.
On June 3, 2015 Christian Kroun Damgaard, Aarhus University wrote:
Western using lysates from HeLa, NHDF, HEK293
1:2000 for western (standard 5% milk, TBS-T buffer).
Secondary antibodies dilution: 1:5.000 of goat anti-rabbit/HRP antibodies in 5%milk/TBS-T (DAKO #P0448) (incubation time: 1hr at RT)
Chemiluminescent substrate: SuperSignal West Femto (Thermo Scientific #34096)
Clean and robust band at approximately 70 kDa detected.
Immunofluorescence using HeLa, NHDF, HEK293 cells fixed in 4% PFA (PBS), Permeabilization in 0.5% Triton/PBS/1% horse serum). Primary: 1:1000 dilution in PBS/1% horse serum. Secondary: 1:1000 dilution PBS/1% horse serum. Donkey anti rabbit – Alexa 488 or Alexa 594 conjugated. Bright cytoplasmic staining.