We performed on-blot biotinylation of sialic acids 53 . We subjected recombinant, purified flagellins to SDS-PAGE and western blotting on to nitrocellulose membrane. We performed on-blot oxidation with 1 mM sodium metaperiodate in phosphate buffered saline (PBS) (30 mM sodium phosphate, pH 7.4 with 150 mM NaCl) at 4°C for 20 minutes. We then washed the blot with PBS and incubated it in PBS, freshly adjusted to pH 6.7 and containing 250 µM amino-oxybiotinoyl-hydrazine (Invitrogen) and 10 mM aniline, for one hour at room temperature in the dark. We followed this up with washing the blot, overnight blocking in 5% skimmed milk, and probing the blot with HRP-conjugated streptavidin (Jackson ImmunoResearch) followed by detection with Luminata Forte western HRP substrate (Merck-Millipore).
Amino oxybiotinoyl hydrazine
Manufactured by Thermo Fisher Scientific
Amino-oxybiotinoyl-hydrazine is a chemical compound used in various laboratory applications. It functions as a reagent for the detection and labeling of biomolecules, such as proteins and nucleic acids. The core functionality of this product is to facilitate the incorporation of biotin, a vitamin, into target molecules, enabling their subsequent identification and analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using amino oxybiotinoyl hydrazine
1
On-Blot Sialic Acid Biotinylation
2
On-Blot Sialic Acid Biotinylation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We performed on-blot biotinylation of sialic acids 53 . We subjected recombinant, purified flagellins to SDS-PAGE and western blotting on to nitrocellulose membrane. We performed on-blot oxidation with 1 mM sodium metaperiodate in phosphate buffered saline (PBS) (30 mM sodium phosphate, pH 7.4 with 150 mM NaCl) at 4°C for 20 minutes. We then washed the blot with PBS and incubated it in PBS, freshly adjusted to pH 6.7 and containing 250 µM amino-oxybiotinoyl-hydrazine (Invitrogen) and 10 mM aniline, for one hour at room temperature in the dark. We followed this up with washing the blot, overnight blocking in 5% skimmed milk, and probing the blot with HRP-conjugated streptavidin (Jackson ImmunoResearch) followed by detection with Luminata Forte western HRP substrate (Merck-Millipore).
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!