The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 240 kDa (predicted molecular weight: 269 kDa).
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Probably involved in connections of major cytoskeletal structures to the plasma membrane. High molecular weight cytoskeletal protein concentrated at regions of cell-substratum contact and, in lymphocytes, at cell-cell contacts.
Contains 1 FERM domain. Contains 1 I/LWEQ domain.
Cell projection, ruffle membrane. Cytoplasm, cytoskeleton. Cell surface. Cell junction, focal adhesion. Colocalizes with LAYN at the membrane ruffles. Localized preferentially in focal adhesions than fibrillar adhesions (By similarity).
Overlay histogram showing RAW 264.7 cells stained with ab157808 (red line). The cells were fixed with 80% methanol (5 min)/ and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab157808, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in RAW 264.7 cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - Anti-Talin 1 antibody [8D4] (ab157808)
Lanes 1 & 3 : PC12 cell lysate - stimulated with calyculin A at 100nM for 30 minutes Lanes 2 & 4 : PC12 cell lysate - stimulated with calyculin A at 100nM for 30 minutes and treated with lambda phosphatase