LC3B Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Dilute Primary Ab dilution to-1:50. - Incubate Ab for overnight at 4°C

Protocol tips
- Dilute Primary Ab dilution to-1:50. - Incubate Ab for overnight at 4°C

Publication protocol

Total proteins were extracted using RIPA buffer containing protease and phosphatase inhibitors for the surgical sciatic nerves or RSC96 cells. The protein concentration was quantitatively analysed, using a BCA protein assay kit (Thermo Fisher Scientific, Rockford, IL, USA). Samples containing 80 μg total protein were separated from 10% or 12% SDS‐polyacrylamide gels and transferred into the nitrocellulose membrane PVDF (Bio‐Rad, Hercules, CA, USA). After blocking with 5% nonfat milk, the membranes were incubated with the following primary antibodies: GFAP (1:10000; Abcam, ab10062), S100 (1:200, Abcam, ab4066), MBP (1:1000; Abcam, ab40390), ERK (1:100; Santa Cruz, sc‐14), p‐ERK (1:1000; Cell Signaling Technology, 4370S), Nrf‐2 (1:1000; Abcam, ab62352), HO‐1 (1:100; Santa Cruz, sc‐1797), NQO1 (1:1000; Abcam, ab34173), Bcl‐2 (1:1000; Abcam, ab59348), BAX (1:1000; Cell Signaling Technology, 14796S), Actived‐caspase3 (1:1000; Abcam, ab2302), Beclin‐1 (1:2000; Abcam, B6186), Atg‐5 (1:1000; Cell Signaling Technology, A074), and LC3 (1:50; Novus, NB100‐2220) overnight at 4°C. Then, the membranes were washed with TBST three times and incubated with horseradish peroxidase‐conjugated secondary antibodies (1: 10000) for 1 hour. GAPDH (1:10000; MultiSciences, 70‐Mab5465‐100) was used as an internal control. Signals were visualized and band density was quantified, using a ChemiDocXRS + imaging system (Bio‐Rad Laboratories, Hercules, CA, USA). The above experiment was repeated three times.

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 - RSC96 using LC3B Antibody from Novus Biologicals.

Manufacturer protocol

Download the product protocol from Novus Biologicals for LC3B Antibody below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - RSC96 using LC3B Antibody from Novus Biologicals. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.