Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Breast cancer)
Specification
Breast cancer
Fixative
Methanol-Acetone
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 25°C
Other product details
Dilution
1/200
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS-milk 0.1%
Secondary antibody
Name
Non-Abcam antibody was used: ALEXAFluor™ 594 goat anti rabbit Invitrogen
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Dilution
1/1500
Additional data
Additional Notes
fixation/permeabilization: ice-cold methanol-aceton (1:1) 15 min on ice
wash/block: 3x10min in PBS-milk 0.1%
incubation with primary antibodies: 1:200 in PBS-milk 0,1% 4°C overnight
wash: 3xPBS 10min
incubation with secondary antibodies: AlexaFluor 594 1:1500 in PBS-milk 0,1% 2h at RT
wash:1xPBS 5min
DAPI: 10min RT
wash: 2xPBS 5 min
wash/block: 3x10min in PBS-milk 0.1%
incubation with primary antibodies: 1:200 in PBS-milk 0,1% 4°C overnight
wash: 3xPBS 10min
incubation with secondary antibodies: AlexaFluor 594 1:1500 in PBS-milk 0,1% 2h at RT
wash:1xPBS 5min
DAPI: 10min RT
wash: 2xPBS 5 min
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投稿 Aug 07 2012