Specificityab78291 reacts with an N-terminal epitope in human Talin 1, between amino acids 139-433. This antibody 100% homologous to human Talin 1 and 85% homologous to human Talin 2. Is is likely that this antibody will also detect Talin 2. On microinjection into whole cells ab78291 disrupts actin stress fibers and focal contacts and inhibits cell motility.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use at an assay dependent concentration. Predicted molecular weight: 270 kDa.
Use at an assay dependent concentration.
Use a concentration of 5 µg/ml.
FunctionProbably 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.
Sequence similaritiesContains 1 FERM domain. Contains 1 I/LWEQ domain.
Cellular localizationCell 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).
ICC/IF image of ab78291 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab78291 at 5 µg/mL overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti-Mouse IgG (H+L) (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of Talin 1 staining in human muscle formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78291, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
References for Anti-Talin 1 antibody [TA205] (ab78291)
This product has been referenced in:
Bolton, S.J. , et al. Monoclonal antibodies recognizing the N- and C-terminal regions of talin disrupt actin stress fibers when microinjected into human fibroblasts Cell Motility and the Cytoskeleton36:363 - 376 (1997).
WB, IF, IP
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