PABPN1, Abcam, ab75855

Very good mAb for IF (4% PFA fixed HeLa cells).
1:2000 dilution in 2% BSA/PBS, 1 h room temperature incubation.

Extremely sensitive antibody. Provides very specific detection of protein with no additional background (in tested conditions). Used for Western Blot detection of PABPN1 protein in whole cell extract from HeLa Kyoto line.

Experimental details:

-primary antibodies dilution: 1:100.000 of PABPN1 antibody in 5%milk/PBS-T (incubation time: 1hr at RT),

-secondary antibodies dilution: 1:10.000 of goat anti-rabbit/HRP antibodies in 5%milk/PBS-T (DAKO #P0448) (incubation time: 1hr at RT),

-chemiluminescent substrate: SuperSignal West Femto (Thermo Scientific #34096)

– amount of loaded material: 5µg of whole cell extract was loaded per lane (measured by Bradford)

– gel system: 4-12% Bis-Tris gel (1xMOPS running buffer, at 200V).

Protein was detected just below ~55kDa band of PageRuler Plus Prestain Protein Ladder (Thermo Scientific #26619).