A549, H1650 and H226 cells were fixed in 4% paraformaldehyde (Boston Bioproducts), permeabilized in cold methanol (Fisher), and immunofluorescence staining was performed using an antibody against NF-κB2 p100/52 (Cell Signaling Technology; Cat no.:3017). The cells were then washed with PBS and incubated with Alexa Fluor 568-conjugated anti-rabbit secondary antibody (Molecular Probes; Cat no.:A11036). All images were captured by confocal microscopy (Nikon; 60x objective, 1.27 NA) and processed using ImageJ software (Version 1.6.0_24; https://imagej.nih.gov/ij ). DAPI-stained nuclei were segmented manually by drawing a circular ROI. Nuclear colocalization of NF-κB2 p52 (red fluorescent signal) with DAPI was measured. The mean nuclear fluorescence was calculated and plotted in GraphPad Prism software (Version 7).
Antibody against nf κb2 p100 52
Manufactured by Cell Signaling Technology
Antibody against NF-κB2 p100/52 is a tool used for the detection and analysis of the NF-κB2 protein, a member of the NF-κB transcription factor family. It recognizes both the precursor form (p100) and the mature form (p52) of the NF-κB2 subunit.
Automatically generated - may contain errors
2 protocols using antibody against nf κb2 p100 52
1
Quantifying NF-κB2 p52 Localization
2
Quantifying Nuclear NF-κB2 Localization
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A549, H1650 and H226 cells were fixed in 4% paraformaldehyde (Boston Bioproducts), permeabilized in cold methanol (Fisher) and immunofluorescence staining was performed using an antibody against NF-κB2 p100/52 (Cell Signaling Technology; cat no.: 3017). The cells were then washed with PBS and incubated with Alexa Fluor 568-conjugated anti-rabbit secondary antibody (Molecular Probes; cat no.: A11036). All images were captured by confocal microscopy (Nikon; 60× objective, 1.27 NA) and processed using ImageJ software (Version 1.6.0_24; https://imagej.nih.gov/ij ). The 4',6-diamidino-2-phenylindole (DAPI)-stained nuclei were segmented manually by drawing a circular region of interest. Nuclear colocalization of NF-κB2 p52 (red fluorescent signal) with DAPI was measured. The mean nuclear fluorescence was calculated and plotted in GraphPad Prism software (Version 7).
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