Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb

Autophagy assay cell type - THP 1

Experiment
Autophagy assay cell type - THP 1
Product
Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb from Cell Signaling Technology
Manufacturer
Cell Signaling Technology

Protocol tips

Upstream tips
Lyse cells in in a modified RIPA 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
Dilute primary Ab at 1:1,000 and incubate 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

Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb from Cell Signaling Technology has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - THP 1 using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb from Cell Signaling Technology.

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

Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - THP 1 using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb from Cell Signaling Technology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms