Anti-p62 (SQSTM1) (Human) pAb

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Lyse cells in RIPA buffer (50 mM Tris, 1.0 mM EDTA, 150 mM NaCl, 0.1% Triton X-100, 1% sodium deoxycholate, 1 mM PMSF)	- Incubate primary antibody at 4 °C overnight

Upstream tips
- Lyse cells in RIPA buffer (50 mM Tris, 1.0 mM EDTA, 150 mM NaCl, 0.1% Triton X-100, 1% sodium deoxycholate, 1 mM PMSF)
Protocol tips
- Incubate primary antibody at 4 °C overnight

Publication protocol

Western blotting
Cells were lysed with RIPA buffer (50 mM Tris, 1.0 mM EDTA, 150 mM NaCl, 0.1% Triton X-100, 1% sodium deoxycholate, 1 mM PMSF). Protein concentrations were quantified by the BCA protein assay kit (Pierce, 23225). Proteins were resolved on 12% SDS-PAGE, then transferred to PVDF (Bio-Rad) membranes. After blocking, the membranes were incubated with primary antibodies at 4 °C overnight, then incubated with secondary antibodies at room temperature for 1 h. Target proteins were examined using Enhanced Chemiluminescence reagents (Millipore, WBKLS0100).

Immunofluorescence
Cells were fixed with 4% paraformaldehyde for 30 min, washed with PBS, and then incubated with 0.1% Triton X-100 for permeabilization. Cells were stained with anti-LC3B polyclonal antibody at 4 °C overnight, and subsequently with Alexa Fluor 488-conjugated goat anti-rabbit LgG (Abcam, ab150077) at 37 °C for 1 h. Nuclei were stained with DAPI for 5 min. Images were captured using a confocal laser scanning microscope (Olympus, FV3000).

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 - A549 using Anti-p62 (SQSTM1) (Human) pAb from MBL international corporation.

Manufacturer protocol

Download the product protocol from MBL international corporation for Anti-p62 (SQSTM1) (Human) pAb below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - A549 using Anti-p62 (SQSTM1) (Human) pAb from MBL international corporation. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.