Purified SFB fractions were labeled at RT with 2 mM sulfosuccinimidyl biotin NHS-LC-LC-Biotin (ThermoFisher, 21343) in PBS for 45 min, quenched with 500 mM glycine, washed twice in PBS and frozen at -80°C. Thawed samples were lysed in PBS with protease inhibitors (Roche, Complete Mini) by sonication (5x30 sec rounds, 30 Khz sonicator at 80%, Hielscher UP50H), and ultracentrifuged at 100,000xg for one hour at 4°C. Pellets were resuspended in CHAPS(4%)/ASB-14(4%) (Sigma), dissociated by 3x20 sec rounds with the FastPrep dissociator (MPBio, 0.1 mm beads), and incubated for 20 min on ice. The streptavidin bead purification was performed as per vendor’s manual (Dynabeads MyOne Streptavidin C1, ThermoFisher). Proteins were eluted at 95°C for 5 min in 1x Laemmli and digested in S-Trap™ micro spin columns (Protifi) as per vendor’s protocol. After elution, peptides were injected in a nanoRSLC-Q Exactive PLUS (RSLC Ultimate 3000) (ThermoScientific) as previously described (PMID:31042281). MS files were processed with Proteome Discoverer software (v1.4) and searched with Mascot search engine against SFB proteins.
Chaps 4 asb 14 4
Manufactured by Merck Group
CHAPS(4%)/ASB-14(4%) is a laboratory reagent used for protein solubilization and extraction. It is a mixture of two zwitterionic detergents, CHAPS (4%) and ASB-14 (4%), which are commonly used in various biochemical applications, such as protein purification and sample preparation for analysis.
Automatically generated - may contain errors
Lab products found in correlation
Dynabeads myone streptavidin c1, by Thermo Fisher Scientific (2 mentions)
Nanorslc q exactive plus rslc ultimate 3000, by Thermo Fisher Scientific (2 mentions)
Nhs lc lc biotin, by Thermo Fisher Scientific (2 mentions)
S trap micro spin column, by Protifi (2 mentions)
Complete mini, by Roche (2 mentions)
2 protocols using chaps 4 asb 14 4
1
Purification and Identification of SFB Proteins
2
Purification and Identification of SFB Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Purified SFB fractions were labeled at RT with 2 mM sulfosuccinimidyl biotin NHS-LC-LC-Biotin (ThermoFisher, 21343) in PBS for 45 min, quenched with 500 mM glycine, washed twice in PBS and frozen at -80°C. Thawed samples were lysed in PBS with protease inhibitors (Roche, Complete Mini) by sonication (5x30 sec rounds, 30 Khz sonicator at 80%, Hielscher UP50H), and ultracentrifuged at 100,000xg for one hour at 4°C. Pellets were resuspended in CHAPS(4%)/ASB-14(4%) (Sigma), dissociated by 3x20 sec rounds with the FastPrep dissociator (MPBio, 0.1 mm beads), and incubated for 20 min on ice. The streptavidin bead purification was performed as per vendor’s manual (Dynabeads MyOne Streptavidin C1, ThermoFisher). Proteins were eluted at 95°C for 5 min in 1x Laemmli and digested in S-Trap™ micro spin columns (Protifi) as per vendor’s protocol. After elution, peptides were injected in a nanoRSLC-Q Exactive PLUS (RSLC Ultimate 3000) (ThermoScientific) as previously described (PMID:31042281). MS files were processed with Proteome Discoverer software (v1.4) and searched with Mascot search engine against SFB proteins.
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!