Recombinant Anti-PINCH 1 + PINCH 2 antibody [EP1944] (ab108609)

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling PINCH 1 + PINCH 2 with ab108609 at 1/1000 dilution. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling PINCH 1 + PINCH 2 with ab108609 at 1/50 dilution, followed by AlexaFluor® 488-conjugated Goat anti-Rabbit secondary antibody (ab150081) at 1/1000 dilution at RT for 45 min. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594), used as a counterstain at 1/200 dilution, was co-incubated with ab108609 overnight at 4° C. Nucleus were visualized using DAPI.

293T cells were fixed with 100μl 4% PFA for 10 min at RT, then permeabilized with 100μl 90% methanol for 30 min (-20° C) followed by blocking with 10% goat serum for 1h at room temperature. Incubate ab108609 for 30 min at room temperature (RT) and then incubate Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) for 30 min at room temperature.