Anti-YY1 antibody (ab43058)

  Western blot - YY1 antibody (ab43058)

Anti-YY1 antibody (ab43058) at 2 µg/ml + Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate (ab14844) at 20 µg

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 45 kDa
Observed band size : 58 kDa (why is the actual band size different from the predicted?)
Additional bands at : 85 kDa. We are unsure as to the identity of these extra bands.

The band observed at 58 kDa is consistent with the banding pattern seen in other commercially available antibodies to YY1.

  Immunocytochemistry/ Immunofluorescence - YY1 antibody (ab43058)

ICC/IF image of ab43058 stained human HEK 293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab43058, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HepG2 and MCF7 cells.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - YY1 antibody (ab43058)

IHC image of YY1 staining in human breast carcinoma FFPE section, performed on a Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab43058, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.