DEF Antibody (N-term) (Cat. #AP11810a)immunohistochemistry analysis in formalin fixed and paraffin embedded human hepatocarcinoma followed by peroxidase conjugation of the secondary antibody and DAB staining.This data demonstrates the use of DEF Antibody (N-term) for immunohistochemistry. Clinical relevance has not been evaluated.
Confocal immunofluorescent analysis of DEF Antibody (N-term)(Cat#AP11810a) with Hela cell followed by Alexa Fluor 488-conjugated goat anti-rabbit lgG (green). DAPI was used to stain the cell nuclear (blue).
Overlay histogram showing U-2OS cells stained with AP11810a (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP11810a, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(OH191631) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Anti-DEF Antibody (N-term) at 1:2000 dilution + A431 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 87 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes : Anti-DEF Antibody (N-term) at 1:2000 dilution Lane 1: HT-1080 whole cell lysate Lane 2: K562 whole cell lysate Lane 3: RPMI 8226 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 87 kDa Blocking/Dilution buffer: 5% NFDM/TBST.