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Recombinant fragment containing amino acids in the C terminal region of Mouse E Cadherin. This sequence is highly conserved in human and rat E Cadherin.
Our Abpromise guarantee covers the use of ab76055 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use at an assay dependent concentration. PubMed: 23405249|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 21769484|
|WB||1/100 - 1/1000. Predicted molecular weight: 97 kDa.
This product may give a weak signal in Western Blot when using unstimulated cell lines. Abcam recommends using A431 cells treated with pervandate (1mM, 30 minutes) as a positive control.
ab76055 staining E Cadherin in mouse intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 0.1% goat Fab anti-mouse IgG for 30 minutes at 25°C; antigen retrieval was by heat mediation in 10mM citrate buffer, pH 6. Samples were incubated with primary antibody (1/250) for 2 hours at 25°C. An Alexa Fluor® 488-conjugated donkey anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.
Blocked and probed in the presence of 5% milk, and the image was captured using chemiluminescence and film.
ICC/IF image of ab76055 stained A431 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 (ab76055, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) 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.
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