Anti-alpha Adaptin抗体[AC1-M11] (ab2807)

Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with Pierce™ Power Blotter System (22834). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.

Immunohistochemistry was performed on normal biopsies of deparaffinized Human heart tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Mouse monoclonal antibody recognizing alpha Adaptin (ab2807) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunolocalization of alpha-adaptin in NRK cells using ab2807 (low power image of image 2).