HEK293 cells were grown in 6-well gelatinized cell culture plates and transfected with 2 μg of the expression constructs per well using Lipofectamine 2000 (Life Technologies), according to the manufacturer's protocol. On the next day after transfections, cells were treated with 5 μM all-trans-retinol for 8 h, or 5 μM all-trans-retinaldehyde for 4 h. Cells were harvested, and retinoids were extracted, separated using normal-phase HPLC and quantified as described before [3 (link),30 (link)]. The expression of recombinant proteins in cell homogenates was detected by Western blotting with anti-FLAG epitope antibody (M7425, Sigma Aldrich) at 1: 2500 dilution.
Anti flag epitope antibody
Manufactured by Merck Group
The Anti-FLAG epitope antibody is a laboratory tool used to detect and purify proteins tagged with the FLAG peptide sequence. It recognizes and binds to this specific epitope tag, allowing researchers to identify and isolate the target protein for further analysis.
Automatically generated - may contain errors
2 protocols using anti flag epitope antibody
1
Retinoid Quantification in Transfected HEK293 Cells
2
Antibody-based Akt Signaling Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies against Akt, phospho-Akt (Ser 473/4 and Thr 308/9- RRIDs- AB_329825 and AB_2255933), phospho-AS160 (Thr642), were obtained from Cell Signaling Technologies. Anti-Flag epitope antibody was purchased from Sigma-Aldrich. Anti-AS160 antibody was purchased from Millipore and anti-actin antibody was purchased from Cytoskeleton Inc. Anti-HA epitope antibody was purchased from Covance (RRID:AB_2565006 ). Cy3 labeled secondary antibody was obtained from Jackson ImmunoResearch Laboratories. Akt inhibitor MK-2206 was purchased from Cayman.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!