This is a knockout-validated antibody summary, based on the publication "Viral FGARAT Homolog ORF75 of Rhesus Monkey Rhadinovirus Effects Proteasomal Degradation of the ND10 Components SP100 and PML", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG2b (kappa light chain)

Company: Santa Cruz

Antibody: PML

Catalog number: sc-377340

Summary: Mouse monoclonal IgG2b (kappa light chain) against amino acids 157-394 of human PML. Recommended for detection of all isoforms of PML of human origin by western blot, immunoprecipitation, immunofluorescence, immunohistochemistry (paraffin) and ELISA.

Western blot

WT and PML-knockout SLK cells. Cells were lysed in RIPA buffer (1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 150 mM NaCl, 1 mM EDTA, 50 mM Tris, pH 8, to which aprotinin and leupeptin were freshly added) for 20 min on ice.

PBS-T supplemented with 5% dry milk powder for at least 30 min.

1:500 dilution in NET gelatin buffer (150 mM NaCl, 5 mM EDTA, 50 mM Tris, 0.05% Triton X-100, 0.25% gelatin) with 0.05% sodium azide or in PBS supplemented with 0.05% Tween (PBS-T) and 5% dry milk powder.

1:2,000 dilution horseradish peroxidase-conjugated secondary antibodies (Dako) and Alexa Fluor 647-conjugated secondary antibodies (Life Technologies) in PBS-T supplemented with 5% dry milk powder.

See Figure 4a in the article [1]. Please note the minor nonspecific bands.