Anti smurf2

Manufactured by Abcam

Sourced in United States

Anti-Smurf2 is a laboratory reagent that can be used to detect and quantify the Smurf2 protein, which is involved in the regulation of various cellular processes. This product is intended for research use only and its performance characteristics have not been established for diagnostic or clinical procedures.

Automatically generated - may contain errors

Lab products found in correlation

3 protocols using anti smurf2

Quantifying Protein Levels in MEFs

Check if the same lab product or an alternative is used in the 5 most similar protocols

MEFs grown on six wells were harvested in lysis buffer containing 25 mM Tris-HCl, pH 7.5, 300 mM NaCl and 1% Triton with protease and phosphatase inhibitors. Tissues were minced by a Dounce homogenizer using 1-2 ml RIPA buffer. A total of 30 µg of protein were separated on 10% SDS-PAGE and transferred to PVDF membranes. After blocking of the membranes by 5% dry milk/TTBS, membranes were incubated in following primary antibody solutions: anti-Smurf1 (Novus, 1D7); anti-Smurf2 (Abcam, EP629Y3); anti-Smad1 (Cell Signaling, 9743); anti-Smad2 (Abcam, EP784Y); anti-Smad3 (Abcam, ab28379), anti-Smad5 (Abcam, EP619Y), anti-phospho-Smad1/5 (Cell Signaling, 41D10), anti-phospho-Smad2 (Cell Signaling, 138D4); anti-phospho-Smad3 (Rockland, 600-401-919), GAPDH (Santa Cruz, 0411), HSC70 (Santa Cruz, B-6). Protein detection was carried out using HRP-coupled species-specific secondary antibodies and ECL solution, exposed to Hyperfilm ECL.

Tang Y., Tang L.Y., Xu X., Li C., Deng C, & Zhang Y.E. (2018). Generation of Smurf2 Conditional Knockout Mice. International Journal of Biological Sciences, 14(5), 542-548.

+ Open protocol

+ Expand

Regulation of PPARγ by Smurf E3 Ligases

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

AML12 cells (ATCC CRL-2254) were cultured in DMEM/F12 supplemented with 10% fetal bovine serum (FBS), 0.005 mg/mL insulin, 0.005 mg/mL transferrin, 5 ng/mL selenium, and 40 ng/mL dexamethasone. Hep3B cells were cultured in MEM supplemented with 1% Non-Essential Amino Acids (NEAA) and 10% FBS. Smurf1KO MEFs were cultured in DMEM supplemented with 10% FBS. Primary hepatocytes were isolated by a two-step collagenase perfusion of the liver and cultured as described [45 (link)]. Flag-tagged PPARγ1, PPARγ2 plasmids, and PPRE-Luc reporter plasmids were obtained from Addgene. Flag-tagged PPARγ2ΔPY plasmid was generated using Site Directed Mutagenesis Kit (Agilent Technologies). Myc-tagged Smurf1, Smurf2, and Smurf1CA mutant, HA-tagged Ubiquitin plasmids were described before [13 (link), 18 (link), 46 (link)]. Anti-Smurf1 (Novus, 1D7); anti-Smurf2 (Abcam, EP629Y3); anti-PPARγ (Santa Cruz, sc-7273); anti-PPARα (Rockland, 600-401-4215); anti-PPARδ (ThermoFisher, PA1-823A); anti-HSC70 (Santa Cruz, B-6); Anti-Flag-Peroxidase (A8592, Sigma); anti-HA (Covance, HA11); and anti-Myc (Santa Cruz, 9E10) were used for western blotting and immunoprecipitation. Knockdown experiments were performed using the following siRNAs: siPPARγ (J-040712-05 and J-040712-07, Dharmacon). Validated siSmurf1, siSmurf2 and siNS were previously described [47 (link), 48 (link)].

Zhu K., Tang Y., Xu X., Dang H., Tang L.Y., Wang X., Wang X.W, & Zhang Y.E. (2018). Non-proteolytic ubiquitin modification of PPARγ by Smurf1 protects the liver from steatosis. PLoS Biology, 16(12), e3000091.

+ Open protocol

+ Expand

Western Blotting and Cell Migration Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis. Western blot analysis was performed as described before (27) . Briefly, after incubation with primary antibody anti-SMURF2 (1:250; Abcam, Cambridge, MA, USA), anti-E-cadherin (1:500; Abcam), anti-N-cadherin (1:200; Abcam), anti-vimentin (1:500; Abcam), anti-SNAI1 (1:500; Abcam), anti-TGFB1 (1:800; Abcam), anti-TWIST (1:500; Abcam), anti-ZEB1 (1:500; Abcam), anti-TGFB2 (1:500; Abcam), anti-α-SMA (1:500; Abcam) and anti-β-actin (1:500; Abcam) overnight at 4˚C, IRDye™-800 conjugated anti-rabbit secondary antibodies (Li-COR, Biosciences, Lincoln, NE, USA) were used for 30 min at room temperature. The specific proteins were visualized by Odyssey™ Infrared Imaging System (Gene Co., Lincoln, NE, USA).
Migration and invasion assay. For transwell migration assays, 5x10 4 cells were plated in the top chamber with the non-coated membrane (24-well insert; pore size, 8 mm; BD Biosciences, San Jose, CA, USA). For invasion assays, 1.25x10

Zhang W.L., Zhang J.H., Wu X.Z., Yan T, & Lv W. (2015). miR-15b promotes epithelial-mesenchymal transition by inhibiting SMURF2 in pancreatic cancer. International journal of oncology, 47(3).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!