Synthetic peptide conjugated to KLH derived from within residues 1100 to the C-terminus of Human Vinculin.
(Peptide available as ab74076.)
Our Abpromise guarantee covers the use of ab73412 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 124 kDa).|
|ICC/IF||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF image of ab73412 stained A549 cells. The cells were 4% formaldehyde fixed (10 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab73412 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
ab73412 staining vinculin in Human platelet cells by Flow cytometry.
Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/250 dilution and incubated for 16 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.
Image courtesy of Dr Mahesh Shivananjappa by Abreview.Blocked with 5% milk for 1 hour at 22°C.