n - biotinyl -1,2 Thermo Fisher Search Results


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    Thermo Fisher biotinylated dhpe
    Saposin B activates and delivers both glycolipid and phospholipids to CD1d in vitro . ( A and B ) In vitro loading of biotin-α-GalCer ( A ) and <t>biotin-DHPE</t> ( B ) on to purified CD1d protein. Soluble CD1d was incubated with <t>biotinylated</t> lipids, and the resulting lipid-CD1d complex was isolated by using neutravidin-agarose beads. Bound CD1d heavy chains were detected by immunoblotting with the D5 mAb. Lanes 1: soluble CD1d used in the loading reaction; lanes 3–11: biotinylated lipids and CHAPS, saposin B, or saposin C added as indicated. ( C and D ) Saposin B does not remove CD1d-associated lipids. The samples shown in lanes 2 and 3 were prepared as in A and B . In lanes 4–9, CD1d was preloaded with biotinylated α-GalCer ( C ) or DHPE ( D ) in the presence of 0.5% CHAPS as in A and B . The neutravidin-agarose beads with associated CD1d–lipid complexes were then incubated under the conditions indicated before analysis of the residual biotinylated lipid-associated CD1d molecules by immunoblotting.
    Biotinylated Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 6 article reviews
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    biotinylated dhpe - by Bioz Stars, 2020-02
    90/100 stars
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    Saposin B activates and delivers both glycolipid and phospholipids to CD1d in vitro . ( A and B ) In vitro loading of biotin-α-GalCer ( A ) and biotin-DHPE ( B ) on to purified CD1d protein. Soluble CD1d was incubated with biotinylated lipids, and the resulting lipid-CD1d complex was isolated by using neutravidin-agarose beads. Bound CD1d heavy chains were detected by immunoblotting with the D5 mAb. Lanes 1: soluble CD1d used in the loading reaction; lanes 3–11: biotinylated lipids and CHAPS, saposin B, or saposin C added as indicated. ( C and D ) Saposin B does not remove CD1d-associated lipids. The samples shown in lanes 2 and 3 were prepared as in A and B . In lanes 4–9, CD1d was preloaded with biotinylated α-GalCer ( C ) or DHPE ( D ) in the presence of 0.5% CHAPS as in A and B . The neutravidin-agarose beads with associated CD1d–lipid complexes were then incubated under the conditions indicated before analysis of the residual biotinylated lipid-associated CD1d molecules by immunoblotting.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Saposin B is the dominant saposin that facilitates lipid binding to human CD1d molecules

    doi: 10.1073/pnas.0700617104

    Figure Lengend Snippet: Saposin B activates and delivers both glycolipid and phospholipids to CD1d in vitro . ( A and B ) In vitro loading of biotin-α-GalCer ( A ) and biotin-DHPE ( B ) on to purified CD1d protein. Soluble CD1d was incubated with biotinylated lipids, and the resulting lipid-CD1d complex was isolated by using neutravidin-agarose beads. Bound CD1d heavy chains were detected by immunoblotting with the D5 mAb. Lanes 1: soluble CD1d used in the loading reaction; lanes 3–11: biotinylated lipids and CHAPS, saposin B, or saposin C added as indicated. ( C and D ) Saposin B does not remove CD1d-associated lipids. The samples shown in lanes 2 and 3 were prepared as in A and B . In lanes 4–9, CD1d was preloaded with biotinylated α-GalCer ( C ) or DHPE ( D ) in the presence of 0.5% CHAPS as in A and B . The neutravidin-agarose beads with associated CD1d–lipid complexes were then incubated under the conditions indicated before analysis of the residual biotinylated lipid-associated CD1d molecules by immunoblotting.

    Article Snippet: Biotinylated DHPE (Molecular Probes) or biotinylated α-GalCer, (2S, 3S, 4R)-1- O -(α- d -galactopyranosyl)-2-[ N -2-{2-[2-(2-biotinylamino-ethoxy)-ethoxy]-ethoxymethoxy}-tetracosanoylamino]-1,3,4-octadecantriol, was sonicated in TBS (pH 5.5) in the presence or absence of detergent or saposins at the indicated concentrations, and soluble CD1d, purified as previously described , was added.

    Techniques: In Vitro, Purification, Incubation, Isolation