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DYDYLNDWGPRFKKLADMYGGGDDconjugated to KLH, corresponding to amino acids 889-912 of Chicken N-Cadherin (cdh2).
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.|
|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.|
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.
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.
This image was kindly supplied by Dr Vladimir Milenkovic by AbreviewGel running conditions are denaturing and 10% SDS.
This image is courtesy of an Abreview submitted by Dr. Vladimir Milenkovic
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.