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This product is useful for the detection of members of the cadherin family or genetically engineered proteins containing the C-terminal cadherin tail, and for demonstration of adherens type cell-cell junctions regardless of their cadherin type.
Our Abpromise guarantee covers the use of ab6528 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1-2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||1/1000. Predicted molecular weight: 125-140 kDa.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
Cellular localization: cell membrane. Specificity This antibody reacts with pan cadherin with a distinct 135 kDa band from a wide variety of tissues. Cadherins are members of a multigene family of single chain glycoprotein receptors mediating Ca2+ dependent cell-cell adhesion
This image is courtesy of an Abreview submitted by Dr. Vladimir Milenkovic
Immunofluorescent imaging of human cells (U2OS) with ab6528
reveals the expected highly specific membranous distribution.
IF was performed with a standard paraformaldehyde technique (fixed in
PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS
for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.
Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour,
secondary antibody for 30 minutes. All blocking and incubation steps
carried out at 37 degrees C.
Overlay histogram showing HeLa cells stained with ab6528 (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 (ab6528, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This image was kindly supplied by Dr Vladimir Milenkovic by AbreviewGel running conditions are denaturing and 10% SDS.Blocked using 5% milk for 30 minutes at 25°C.
ab6528 staining pan Cadherin in mouse Cor1 cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with formaldehyde, permeabilized with TBS, BSA, azide and Triton and blocking with 1% BSA for 30 minutes at RT was performed. Samples were incubated with primary antibody (1/1500: in TBS, BSA, azide and 0.3%Triton) for 2 hours. An Alexa Fluor®546-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/1000 dilution. The image was captured using Z-stacks/Zeiss Apotome microscope rig.
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