Total protein expression of the OR51E1 constructs in HEK 293T cells was measured by enzyme-linked immunosorbent assay (ELISA), as previously reported (34 (link),66 (link)). Cells were grown in a poly-l -lysine 96-well-plate (black plate with clear bottoms) and transfected with OR51E1 or OR51E1 variants with RTP1S and with or without Golf. After 24-hour transfection, cells were fixed, permeabilized, and incubated with mouse monoclonal anti-flag antibody (M2, catalog no. F1802, MilliporeSigma) at 1:100 dilution and detected with anti-mouse horseradish peroxidase (HRP)–conjugated secondary antibody (catalog no. 115-035-146, the Jackson Laboratory) at 1:8000 dilution. HRP levels were measured with 1-Step Ultra TMB (3,3′,5,5′-tetramethylbenzidine; catalog no. 34028, Thermo Fisher Scientific) at excitation of 450 nm.
Protocol detail
Quantifying OR51E1 Protein Expression
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.
Corresponding Organization : Johns Hopkins University
Other organizations : Queen Mary University of London, William Harvey Research Institute
Top 5 similar protocols
Variable analysis
independent variables
- OR51E1 constructs
- Presence of RTP1S
- Presence of Golf
- Total protein expression of OR51E1 constructs in HEK 293T cells
- Cells were grown in a poly-L-lysine 96-well-plate (black plate with clear bottoms)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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.
Sign up for free or login to display all annotations
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!