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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 ab6529 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/1000. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
|WB||1/200. On chicken and rabbit heart extract this antibody detects a single band at 135 kD.|
|ICC/IF||1/100. Fix cells with 2-4% paraformaldehyde for 10 minutes at room temperature, followed by 10-minute incubation with 0.2% Triton X-100 (see Gellersen et al 2007 and Kaur et al 2006).|
|Electron Microscopy||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration. Fix with ice cold ethanol.|
Immunofluorescent imaging of human cells (U2OS) with ab6529 confirms the specificity of this antibody. Pan cadherin antibody recognises predominantly membranous signal with some fainter cytoplasmic staining corresponding to soluble cadherin. No nuclear staining is seen.
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. Nuclei were stained with Hoechst stain.
ab6529 staining pan Cadherin in human bone marrow cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed and blocked in 2% BSA for 30 minutes at 20°C. The primary antibody was diluted, 1/200 and incubated with sample for 9 hour at 4°C. An Alexa Fluor® 488 conjugated goat polyclonal to rabbit IgG, diluted 1/200 was used as secondary.
This image is courtesy of an anonymous AbreviewAll Lanes: pan Cadherin antibody (ab6529) at 1/500 dilution + Mouse brain whole tissue lysate (100 µg)