Non-Abcam antibody was used: Invitrogen Goat Anti Rabbit AF488 Host species: Goat Clonality: Polyclonal Conjugation: Alexa Fluor® 488
Mouse RAW264.7 cell line was grown on cover slips in 6 well plates in DMEM/10% FCS. Cells were washed with PBS prior to fixing with 1%PFA. Wash 3x3min in PBS. Block 10% goat serum for 20min at RT Primary Ab diluted in PBS and incubated overnight at 4deg. Wash 3x3min in PBS. Secondary - goat anti-rabbit AF488 (green) 1/1000 in PBS for 1h at RT. Wash 3x3min in PBS Counterstained with Hoechst 33258 (blue) for 10mins at RT. Wash 3min in dH2O Coverslips removed tapped dry and placed on glass slides and mounted in Invitrogen ProLong gold and kept at 4deg until imaged.
Abcam user community
Submitted Aug 01 2012
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