Anti-SSRP1 antibody (ab21584)

  Western blot - SSRP1 antibody (ab21584)

All lanes : Anti-SSRP1 antibody (ab21584) at 1/500 dilution

Lane 1 : U2OS cell lysate
Lane 2 : U2OS cell lysate treated with ionizing radiation

developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 81 kDa
Observed band size : 90 kDa (why is the actual band size different from the predicted?)
Additional bands at : 70 kDa (possible cleavage fragment,cross reactivity).

From a 60 mm dish, cells were scraped into 75 ul of 2X laemmli buffer and 20ul was loaded onto the SDS-PAGE gel. ab21584 recognizes a protein at approximately 90 kDa which corresponds closely to the size of SSRP1.

  Immunocytochemistry/ Immunofluorescence - SSRP1 antibody (ab21584)

ICC/IF image of ab21584 stained human HEK 293 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab21584, 5µg/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.