SDS-PAGE was performed on OMV preparations (containing approximately 2.5–5 mg/ml protein) using 8–25% gradient gels in the PhastSystem (GE Healthcare, Piscataway, NJ). Molecular weights of the reduced and denatured OMV proteins from the three Cronobacter spp. were estimated by using the relative mobility method (Weber et al., 1972 (link)). Western blotting of OMV protein samples separated by SDS-PAGE was carried out by electro-blotting onto a Problott (Applied Biosystems, Foster City, CA) membrane in a transfer buffer (10 mM of 3-[cyclohexylamino]-1-propane sulfonic acid containing 10% methanol, pH 11).
Problott
Manufactured by Thermo Fisher Scientific
Sourced in United States
ProBlott is a transfer membrane used for Western blotting techniques. It serves as a support for the transfer of proteins from polyacrylamide gels to a solid matrix, allowing for subsequent detection and analysis.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using problott
1
SDS-PAGE and Western Blotting of OMVs
2
SDS-PAGE and Immunoblotting for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were separated by SDS-PAGE in 10 or 12 % gels (45) (link). Gels were either stained with Coomassie Brilliant Blue G-250 (Serva, USA) or electroblotted onto PVDFmembrane (ProBlott, Applied Biosystems, USA) following the manufacturer's instructions. RJ proteins were detected using specific anti-apalbumin pAb as was described previously (5, (link)42, 43) . Immunoactive protein bands were visualised by incubating the blots in the chromogenic DAB/ NiCl 2 solution.
3
Western Blot Analysis of Apolipoproteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The isolated RLP sample or plasma was subjected to 12% SDSpolyacrylamide gel electrophoresis and the proteins in the gel were transferred to a polyvinylidene difluoride membrane (ProBlott; Applied Biosystems) by electroblot. The membranes were blocked with 5% dry milk for 2 h and incubated with primary antibodies for 16 h. The membranes were then incubated with a second antibody conjugated with alkaline phosphatase for 2 h, then visualized using the nitro blue tetrazolium/5-bromo-4-chloro-3-indolylphosphate p-toluidine salt system. Anti-apolipoprotein A-I antibody (600-101-109) was from Rockland and anti-apolipoprotein B-100 antibody (20H-G1b) was from the Academy Bio-Medical Co.
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!