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    Stain-free protein quantification and normalization
    6.
    发明授权
    Stain-free protein quantification and normalization 有权

    公开(公告)号：US09606111B2

    公开(公告)日：2017-03-28

    申请号：US13870710

    申请日：2013-04-25

    Applicant: Bio-Rad Laboratories, Inc., LSG-LSD Division

    Inventor： Steve Freeby ,  Ning Liu ,  Kevin McDonald ,  Aran Paulus ,  Anton Posch

    IPC: G01N33/53 ,  G01N33/68

    CPC classification number: G01N33/53 ,  G01N33/6827 ,  G06F19/10 ,  G06T7/0012 ,  G06T11/60 ,  G06T2207/30004

    Abstract: Disclosed herein are methods of protein quantification and normalization using haloalkylated tryptophan fluorescence. Complex protein samples, i.e., samples that each contain 1,000 or more distinct proteins, from diverse sources that do not have common protein profiles are treated with a halo-substituted organic compound (i.e. haloalkane) that reacts with tryptophan residues to form fluorescent products. Irradiation of the samples with ultraviolet light and the detection and quantification of the resultant fluorescent emissions from all proteins in each sample are then used to obtain comparative values for total protein content among the various samples. The values thus obtained are found to be valid indications of comparative total protein content, despite the fact that the tryptophan levels vary widely among the various proteins in any single sample and the samples, due to the diversity of their origins, tend to differ among themselves in the identities and relative amounts of the proteins that they contain. Protein samples are also normalized to correct for differences in sample dilution, sample loading, and protein transfer inconsistencies, by using stain-free detection of total protein in each of the samples, or detection of subsamples within each sample.

    STAIN-FREE PROTEIN QUANTIFICATION AND NORMALIZATION
    7.
    发明申请
    STAIN-FREE PROTEIN QUANTIFICATION AND NORMALIZATION 审中-公开
    无蛋白质定量和正常化

    公开(公告)号：US20170030922A1

    公开(公告)日：2017-02-02

    申请号：US15293154

    申请日：2016-10-13

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Steve Freeby ,  Ning Liu ,  Kevin McDonald ,  Aran Paulus ,  Anton Posch

    IPC: G01N33/68 ,  G06T7/00 ,  G06T11/60 ,  G06F19/10

    CPC classification number: G01N33/53 ,  G01N33/6827 ,  G06F19/10 ,  G06T7/0012 ,  G06T11/60 ,  G06T2207/30004

    Abstract: Disclosed herein are methods of protein quantification and normalization using haloalkylated tryptophan fluorescence. Complex protein samples, i.e., samples that each contain 1,000 or more distinct proteins, from diverse sources that do not have common protein profiles are treated with a halo-substituted organic compound (i.e. haloalkane) that reacts with tryptophan residues to form fluorescent products. Irradiation of the samples with ultraviolet light and the detection and quantification of the resultant fluorescent emissions from all proteins in each sample are then used to obtain comparative values for total protein content among the various samples. The values thus obtained are found to be valid indications of comparative total protein content, despite the fact that the tryptophan levels vary widely among the various proteins in any single sample and the samples, due to the diversity of their origins, tend to differ among themselves in the identities and relative amounts of the proteins that they contain. Protein samples are also normalized to correct for differences in sample dilution, sample loading, and protein transfer inconsistencies, by using stain-free detection of total protein in each of the samples, or detection of subsamples within each sample.

    Abstract translation: 本文公开了使用卤代烷基化色氨酸荧光进行蛋白质定量和归一化的方法。 使用与色氨酸残基反应以形成荧光产物的卤素取代的有机化合物（即卤代烷）处理复合蛋白质样品，即每种含有来自不同来源的不具有共同蛋白质谱的1,000个或更多个不同蛋白质的样品。 然后使用紫外光照射样品，并对每个样品中所有蛋白质产生的荧光发射进行检测和定量，以获得各种样品中总蛋白质含量的比较值。 这样获得的值被发现是比较总蛋白质含量的有效指示，尽管事实上，任何单个样品中的各种蛋白质之间的色氨酸水平变化很大，并且样品由于其起源的多样性而趋于彼此不同 在它们所含的蛋白质的身份和相对量中。 蛋白质样品也通过使用每个样品中的总蛋白的无染色检测或每个样品中的子样品检测来校准样品稀释度，样品加载和蛋白质转移不一致性的差异。

    Stain-free protein quantification and normalization
    10.
    发明授权
    Stain-free protein quantification and normalization 有权

    公开(公告)号：US10539556B2

    公开(公告)日：2020-01-21

    申请号：US16158853

    申请日：2018-10-12

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Steve Freeby ,  Ning Liu ,  Kevin McDonald ,  Aran Paulus ,  Anton Posch

    IPC: G01N33/53 ,  G16B99/00 ,  G01N33/68 ,  G06T7/00 ,  G06T11/60

    Abstract: Disclosed herein are methods of protein quantification and normalization using haloalkylated tryptophan fluorescence. Complex protein samples, i.e., samples that each contain 1,000 or more distinct proteins, from diverse sources that do not have common protein profiles are treated with a halo-substituted organic compound (i.e. haloalkane) that reacts with tryptophan residues to form fluorescent products. Irradiation of the samples with ultraviolet light and the detection and quantification of the resultant fluorescent emissions from all proteins in each sample are then used to obtain comparative values for total protein content among the various samples. The values thus obtained are found to be valid indications of comparative total protein content, despite the fact that the tryptophan levels vary widely among the various proteins in any single sample and the samples, due to the diversity of their origins, tend to differ among themselves in the identities and relative amounts of the proteins that they contain. Protein samples are also normalized to correct for differences in sample dilution, sample loading, and protein transfer inconsistencies, by using stain-free detection of total protein in each of the samples, or detection of subsamples within each sample.

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