All lanes : Anti-PRMT3 antibody [EPR13279] (ab191562) at 1/10000 dilution

Lane 1 : HepG2 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : 293 cell lysate
Lane 4 : MCF7 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution

Developed using the ECL technique.

Predicted band size: 60 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Immunohistochemical analysis of paraffin-embedded, human cervix carcinoma tissue labeling PRMT3 with ab191562 at a 1/100 dilution. Counter stained with hematoxylin. In the negative control PBS was used instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescence analysis of, paraformaldehyde-fixed, MCF7 cells labeling PRMT3 with ab191562 at a 1/50 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor^® 488) ab150077 was used at a 1/200 dilution. In blue DAPI staining. In the negative controls cells were treated with anti-PRMT3 at a 1/50 dilution as primary antibody and goat anti-mouse IgG (Alexa Fluor®594) at a 1/400 dilution as secondary antibody.

Western blot analysis on immunoprecipitation from 1) 293 cell lysate and 2) PBS, labeling PRMT3 using ab191562 at 1/40 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG at a 1/1500 dilution.