immunocytochemistry antibody customer review for anti β-actin antibody (stj96930)
TRANSCRIPT
Result
“Antibody seems to specifically label β-actin. “ - J. Heller, University College London.
Cell Line: Rat dissociated cortical cul-
ture (14 DIV)
Method of validation: Immunocytochemistry
Primary Antibody: STJ96930 Anti-β-actin anti-
body
Secondary Antibody Donkey anti-mouse CF568
(1:250)
Dilution ratio: 1:100
Protocol
Treatment of materials: Cortical cells were fixed with 3.7% PFA for 12 min at 37°C.
Membrane wash: PBS 2x 5 min, gently rocking at RT ;
Quenching: 0.1% NaBH4 in PBS for 7 min, gently rocking at RT ;
Membrane wash: PBS 1x 2 and 3x 5 min, gently rocking at RT ;
Permeabilization: PBS supplemented with 0.1% triton X-100 (PBS-T) for 10 min, gently rocking at RT ;
Membrane wash: PBS 1x 2 and 3x 5 min, gently rocking at RT ;
Blocking: blocking buffer (PBS supplemented with 3% BSA) for 30 min, gently rocking at RT ;
Primary antibody probing: PBS supplemented with 0.5% BSA overnight at 4°C ;
Membrane wash: PBS 1x 2 and 3x 5 min, gently rocking at RT ;
Secondary antibody probing: Fuorescently-Labelled secondary antibodies diluted in PBS for 1 h, gently rocking at RT;
Membrane wash: PBS 5x 10 min, gently rocking at RT;
Fixation: 4% PFA in PBS for 10 min at RT ;
Membrane wash: PBS 3x 10 min, gently rocking at RT ;
Visualization: Imaging in PBS.
Antibody Customer Review: STJ96930 Anti-β-actin Antibody Antibody Specificity:
Antibody Rating:
Fig.1
The antibody labelling showed staining below the plasma membrane as expected. Some cell protrusions are also visualised .