Anti-Egr1 antibody [EPR15916] (ab194357) at 1/50000 dilution + 293T treated with 20% FBS at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 57 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue labeling Egr1 with ab194357 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Counter stained with hematoxylin.

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

Confocal image showing nuclear and weakly cytoplasmic staining on Hela cells.

Immunocytometry/Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma ) cells labeling Egr1 with ab194357 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei counterstained with DAPI (blue).

For negative control 1, ab194357 (1:200) was used, followed by (ab150120) anti-mouse secondary antibody at 1/1000 dilution. For negative control 2, (ab7291) mouse primary antibody (1/1000) was used followed by (ab150077) anti-rabbit secondary antibody at 1/1000 dilution.