Fatty Acid Metabolism and Protein Synthesis

We obtained fatty acids (FAs) from Nu-Chek Prep, Inc.; radiolabeled S-35 methionine from Perkin-Elmer (cat. no. NEG709A500UC); SCD1 inhibitor (A939572; cat. no. 19123), Nonidet P-40 (NP-40; cat. no. 600009) and Protein Synthesis kit (cat. no. 601100) from Cayman Chemicals; rabbit anti-YB-1 from ProteinTech; mouse anti-β-actin from Millipore Sigma; rabbit anti-SCD1 from Cell Signal; YB-1-myc plasmid from Addgene; non-targeting control siRNA and YB-1 siRNAs from Qiagen; FA-free BSA from Sigma-Aldrich; Protease inhibitor cocktail from Roche; Fetal Calf serum (FCS) and Delipidated fetal calf serum (DFCS) from Gemini Bio. All FAs added into culture media were conjugated to BSA as previously reported (10 (link)).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Jeffords E., Freeman S., Cole B., Root K., Chekouo T., Melvin R.G., Bemis L, & Simmons GE J.r. (2020). Y-box binding protein 1 acts as a negative regulator of stearoyl CoA desaturase 1 in clear cell renal cell carcinoma. Oncology Letters, 20(5), 165.

Publication 2020

mouse anti-β-actin rabbit anti-SCD1 FA-free BSA Protease inhibitor cocktail Fetal Calf serum (FCS)

Actin Cayman Culture media Fatty acids Fetal calf serum Methionine Mouse Np 40 Plasmid Protease inhibitor Protein synthesis Rabbit Signal cell Sirnas

Corresponding Organization :

Other organizations : College of St. Scholastica, University of Minnesota, Duluth, University of Calgary

Top 5 similar protocols

Variable analysis

independent variables

FA type (obtained from Nu-Chek Prep, Inc.)
SCD1 inhibitor (A939572; from Cayman Chemicals)
YB-1-myc plasmid (from Addgene)
YB-1 siRNAs (from Qiagen)

dependent variables

SCD1 protein expression (measured by Western blot using rabbit anti-SCD1 antibody from Cell Signal)
YB-1 protein expression (measured by Western blot using rabbit anti-YB-1 antibody from ProteinTech)
β-actin protein expression (measured by Western blot using mouse anti-β-actin antibody from Millipore Sigma)

control variables

Radiolabeled S-35 methionine (from Perkin-Elmer)
Nonidet P-40 (NP-40; from Cayman Chemicals)
Protein Synthesis kit (from Cayman Chemicals)
FA-free BSA (from Sigma-Aldrich)
Protease inhibitor cocktail (from Roche)
Fetal Calf serum (FCS) and Delipidated fetal calf serum (DFCS) (from Gemini Bio)

positive controls

Non-targeting control siRNA (from Qiagen)

negative controls

No additional treatment or transfection

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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!