Purified Mouse Anti-p62 Ick ligand Clone 3/P62 LCK LIGAND (RUO)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Lyse cells in buffer containing 150 mM NaCl, 1% NP-40, 50 mM Tris-HCl, pH 8, 0.5% deoxycholic acid, 0.1% SDS 1% Triton X-100, protease and phosphatase inhibitors	Incubate primary Ab overnight at 4C.

Upstream tips
Lyse cells in buffer containing 150 mM NaCl, 1% NP-40, 50 mM Tris-HCl, pH 8, 0.5% deoxycholic acid, 0.1% SDS 1% Triton X-100, protease and phosphatase inhibitors
Protocol tips
Incubate primary Ab overnight at 4C.

Publication protocol

1 × 106 cells were washed twice with PBS and lysed in a modified RIPA buffer containing 150 mM NaCl, 1% NP-40 (Calbiochem, 492015), 50 mM Tris-HCl, pH 8, 0.5% deoxycholic acid (SIGMA, D-6750), 0.1% SDS (SERVA, 39575.02) 1% Triton X-100, protease and phosphatase inhibitors (SIGMA, S8830, S6508 and 450022) The lysates were subjected to electrophoresis on 4–12% NuPage Bis-Tris gels (Life Technologies/Novex, NP0323) and transferred to nitrocellulose membranes (GE Healthcare/Amersham, 10600002 Protran). The membranes were then blocked in 1X PBS-0.1% Tween 20 (SIGMA, P1379) containing 3% BSA (SIGMA, A4503) and probed with specific primary antibodies, overnight at 4°C. After several washes in 1X PBS-0.1% Tween 20 the membranes were incubated with appropriate secondary antibodies conjugated to horseradish peroxidase (Santa Cruz Biotechnology, sc-2004, sc-2005 and sc-2768), used at 1:10,000. Finally, the membranes were washed in 1X PBS-0.1% Tween 20 and immunoreactivity was detected using an enhanced chemiluminescence kit (Thermo Scientific, 32209). Densitometric analysis was performed using ImageJ software.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - THP 1 using Purified Mouse Anti-p62 Ick ligand Clone 3/P62 LCK LIGAND (RUO) from BD Biosciences.

Manufacturer protocol

Download the product protocol from BD Biosciences for Purified Mouse Anti-p62 Ick ligand Clone 3/P62 LCK LIGAND (RUO) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - THP 1 using Purified Mouse Anti-p62 Ick ligand Clone 3/P62 LCK LIGAND (RUO) from BD Biosciences. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.