1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Cellular localizationPIAS1: Nucleus speckle. Interaction with CSRP2 may induce a partial redistribution along the cytoskeleton.
PIAS2: Nucleus speckle. Nucleus, PML body. Nucleus. Colocalizes at least partially with promyelocytic leukemia nuclear bodies (PML NBs) (PubMed:22406621). Colocalizes with SUMO1 in nuclear granules (By similarity).
Western blot - Anti-PIAS1 + PIAS2 antibody [EPR2581Y] (ab77231)
All lanes : Anti-PIAS1 + PIAS2 antibody [EPR2581Y] (ab77231) at 1/50000 dilution
Lane 1 : Human PIAS1 full-length recombinant protein with GST-Tag Lane 2 : Human PIAS2-beta recombinant protein fragment with GST-His-Tag Lane 3 : Human PIAS3 recombinant protein fragment with SUMO-His-Tag
Lysates/proteins at 0.015 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling PIAS1 + PIAS2 with purified ab77231 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.