Application

Immunoblotting: 123kD in membrane fractions; MCF-7 membranes positive control. Confluent cells were washed three times with ice-cold PBS and lysed in 50mM HEPES pH 7.5, 150mM NaCl, 1mM EDTA, 10% glycerine, 1% triton X-100, 20mM Na(4)P(2)O(7), 2mM Na-vanadate, 100mM NaF, 2mM PMSF, 20μg/mL leupeptin, 1mM pNPP(para-nitrophenylphosphate), 20μg/mL aprotinin, and 200μM NH(4)-molybdate. Crude extracts were clarified by centrifugation (10,000 x g 10 min), and supernatants were taken for immunoprecipitation. Protein concentration of cleared lysates were determined by BCA protein assay. {Buhring, et al Luekemia 10:106-116, 1996}.



FACS Analysis

Functional blocking of E-cadherin

Does not work well for immunoprecipitation

Original paper, Buhring, H et al Leukemia 10:106-116, 1996.

Optimal working dilutions must be determined by end user.

Anti-E-Cadherin Antibody, clone 67A4 detects level of E-Cadherin & has been published & validated for use in FC & WB.

Research CategoryCell Structure

Research Sub CategoryAdhesion (CAMs)

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Immunogen

T-47D breast carcinoma line

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-1103

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein A purified immunoglobulin. Liquid in 0.02M PBS with NaCl, pH=7.6, 0.1% Sodium azide.

Format: Purified

Specificity

Antibodies against E-cadherin are suitable to characterize epithelial cells. E-Cadherin is involved in adhesion of epithelial cells (mostly homotypic and homophilic). E-cadherin is also expressed on immature erythroid cells. The antibody 67A4 is suitable to study E-cadherin expression by flow cytometry. In addition, it blocks the interaction of E-cadherin molecules and therefore is suitable to study E-cadherin-mediated signal transduction.

Antigen is expressed in non-neural epithelial tissues.

Storage and Stability

Maintain 2-8°C in undiluted aliquots for up to 12 months after date of receipt.