This is a knockout-examined 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).

Rabbit polyclonal IgG

Company: Proteintech

Antibody: SP100

Catalog number: 11377-1-AP

Summary: Rabbit polyclonal IgG against SP100 fusion protein. Reacts with human by western blot, immunofluorescence, immunohistochemistry and ELISA.

Western blot

WT and SP100-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 .

1:1,000 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 notice the nonspecific band.