BMmacs from Dhpsfl/fl and DhpΔmye mice were infected or not with H. pylori for 18 h. Cells were then washed with PBS and methionine-free, cysteine-free DMEM (GIBCO) was added to the cells for 2 h. This medium was then replaced by methionine-free, cysteine-free DMEM supplemented with 50 μM Click-IT homopropargylglycine (Invitrogen) for 4 h. Proteins were extracted using 1% SDS in 50 mM Tris-HCl containing the Protease Inhibitor Cocktail (Set III, Calbiochem) and the Phosphatase Inhibitor Cocktail (Set I, Calbiochem). Proteins (50 μg) were labeled with biotin azide (Invitrogen) using the Click Chemistry Reagents (Invitrogen), precipitated by methanol, resuspended in 50 mM Tris-HCl, and immunoprecipitated using a NOS2 Ab (Millipore) and the PureProteome Protein A Magnetic Bead System (Millipore). The resulting precipitates were used for western blotting using 4plus Streptavidin HRP Label (Biocare Medical).
Click chemistry reagents
Manufactured by Thermo Fisher Scientific
Click Chemistry Reagents are a collection of specialized chemical compounds designed to facilitate covalent bond formation through click chemistry reactions. These reagents enable rapid and selective modification of biomolecules, such as proteins, nucleic acids, and carbohydrates, making them a valuable tool for various applications in life science research and development.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using click chemistry reagents
1
Metabolic Labeling of NOS2 Interactome
2
Metabolic Labeling of NOS2 Interactome
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMmacs from Dhpsfl/fl and DhpΔmye mice were infected or not with H. pylori for 18 h. Cells were then washed with PBS and methionine-free, cysteine-free DMEM (GIBCO) was added to the cells for 2 h. This medium was then replaced by methionine-free, cysteine-free DMEM supplemented with 50 μM Click-IT homopropargylglycine (Invitrogen) for 4 h. Proteins were extracted using 1% SDS in 50 mM Tris-HCl containing the Protease Inhibitor Cocktail (Set III, Calbiochem) and the Phosphatase Inhibitor Cocktail (Set I, Calbiochem). Proteins (50 μg) were labeled with biotin azide (Invitrogen) using the Click Chemistry Reagents (Invitrogen), precipitated by methanol, resuspended in 50 mM Tris-HCl, and immunoprecipitated using a NOS2 Ab (Millipore) and the PureProteome Protein A Magnetic Bead System (Millipore). The resulting precipitates were used for western blotting using 4plus Streptavidin HRP Label (Biocare Medical).
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!