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    Protein detection using modified cyclodextrins
    13.
    发明授权
    Protein detection using modified cyclodextrins 有权
    使用改性环糊精的蛋白质检测

    公开(公告)号：US09285359B2

    公开(公告)日：2016-03-15

    申请号：US14455609

    申请日：2014-08-08

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Clayton T. McKee ,  William Strong ,  Lee Olech ,  Thomas Berkelman ,  Christopher Belisle

    IPC: G01N33/53 ,  G01N33/58 ,  G01N33/68 ,  G01N27/447

    CPC classification number: G01N33/5306 ,  G01N27/44747 ,  G01N33/582 ,  G01N33/6803 ,  G01N2400/18

    Abstract: A method is provided for detecting a protein using a cyclodextrin covalently linked to at least one label. The cyclodextrin can associate with the protein by sequestering an aromatic amino acid side-chain of the protein in its hydrophobic cavity. After contacting the protein with the cyclodextrin, the label can be detected directly or can undergo a chemical interaction with a reagent to form a detectable product. The label can include an indole moiety, which can react with a halo-substituted organic compound upon exposure to UV light and thereby be rendered fluorescent. Alternatively, the label can include a biotin moiety, which can bind to a binding partner such as avidin, or variants thereof, to form a detectable molecular complex. A labeled cyclodextrin can be used in the present methods to detect a protein of interest in an electrophoresis gel or on a blotting membrane. Aromatic amino acid residues of the protein, in particular tryptophan, remain protected from chemical modification due to sequestration by the cyclodextrin, making these methods compatible with downstream applications that require intact protein. Also provided herein are compositions, kits, and electrophoresis gels for use in detecting proteins.

    Abstract translation: 提供了一种使用共价连接至少一个标签的环糊精来检测蛋白质的方法。 环糊精可以通过在其疏水腔中螯合蛋白质的芳香族氨基酸侧链而与蛋白质结合。 将蛋白质与环糊精接触后，可以直接检测标记物，或者可以与试剂进行化学相互作用以形成可检测的产物。 标记可以包括吲哚部分，其可以在暴露于紫外光时与卤素取代的有机化合物反应，从而变成荧光。 或者，标记可以包括生物素部分，其可以结合结合配偶体，例如抗生物素蛋白或其变体，以形成可检测的分子复合物。 标记的环糊精可用于本方法中以检测电泳凝胶或印迹膜上的目的蛋白质。 蛋白质，特别是色氨酸的芳香族氨基酸残基由于环糊精的螯合而保护免受化学修饰，使得这些方法与需要完整蛋白质的下游应用相容。 本文还提供了用于检测蛋白质的组合物，试剂盒和电泳凝胶。

    Modified electrode buffers for stain-free protein detection in electrophoresis
    14.
    发明授权
    Modified electrode buffers for stain-free protein detection in electrophoresis 有权
    用于电泳中无污染蛋白检测的修饰电极缓冲液

    公开(公告)号：US09005418B2

    公开(公告)日：2015-04-14

    申请号：US13961626

    申请日：2013-08-07

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Christopher Belisle

    IPC: G01N27/447

    CPC classification number: G01N27/44747 ,  G01N27/44713 ,  G01N27/44721 ,  G01N27/44726 ,  G01N27/44739

    Abstract: Proteins that are electrophoretically separated in a gel are derivatized to produce fluorescent emissions by incorporating halo-substituted organic compounds into one or both of the electrode buffer solutions at the two ends of the gel. The halo-substituted compounds used are ones that bear an electric charge at the pH of the buffer solutions and gel, and the polarity of the charge on the compounds is such that the compounds migrate from the electrode buffer into the gel under the electrophoretic influence concurrently with the migration of the proteins into the gel. Once the proteins are separated and distributed within the gel and the gel is fully penetrated with the halo-substituted compounds, the gel is irradiated with ultraviolet light to induce a reaction between the halo-substituted compounds and the proteins through the tryptophan residues on the proteins, producing fluorescent reaction products.

    Abstract translation: 在凝胶中电泳分离的蛋白质通过将卤素取代的有机化合物引入凝胶两端的一个或两个电极缓冲溶液中衍生而产生荧光发射。 使用的卤素取代的化合物是在缓冲溶液和凝胶的pH下具有电荷的化合物，并且化合物上的电荷的极性使得化合物在电泳影响下从电极缓冲液迁移到凝胶中 随着蛋白质迁移到凝胶中。 一旦蛋白质被分离并分布在凝胶中并且凝胶被卤素取代的化合物完全渗透，则用紫外线照射凝胶以通过蛋白质上的色氨酸残基诱导卤素取代的化合物与蛋白质之间的反应 ，产生荧光反应产物。

    Dynamic axial compression for preparative columns using external compression
    16.
    发明授权
    Dynamic axial compression for preparative columns using external compression 有权

    公开(公告)号：US11841350B2

    公开(公告)日：2023-12-12

    申请号：US17695672

    申请日：2022-03-15

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Christopher Belisle ,  Mark A. Snyder

    IPC: B01D15/22 ,  G01N30/52 ,  G01N30/60

    CPC classification number: G01N30/52 ,  B01D15/22 ,  G01N30/6021 ,  G01N2030/522

    Abstract: A dynamic axial compression column is disclosed herein. This dynamic axial column utilized external compression to prevent the creation of end plate space in the column. The dynamic axial column can include a tube defining a first opening, a second opening, and a lumen extending there between. The dynamic axial column can include a first end plate assembly sealing the first opening and movably extending at least partially into the lumen via the first opening, a second end plate assembly sealing the second opening, a plurality of rods extending along the outside of the tube and connecting the first end plate assembly and the second end plate assembly, and a first plurality of compression devices external to the tube and engaging one of the plurality of rods to bias the first end plate assembly towards the second end plate assembly.

    GeLC-MS using stain free technology
    17.
    发明授权
    GeLC-MS using stain free technology 有权
    GeLC-MS使用无污染的技术

    公开(公告)号：US09488616B2

    公开(公告)日：2016-11-08

    申请号：US13896572

    申请日：2013-05-17

    Applicant: Bio-Rad Laboratories, Inc.

    Inventor： Christopher Belisle

    IPC: G01N27/447 ,  H01J49/00 ,  G01N30/06 ,  G01N33/68 ,  H01J49/26

    CPC classification number: G01N27/44721 ,  G01N30/06 ,  G01N33/6848 ,  H01J49/0036 ,  H01J49/26

    Abstract: Disclosed herein is a method of preparing a protein sample for mass spectroscopy. The method includes separating proteins of the sample on an electrophoresis gel; contacting the proteins with a halo-substituted organic compound; exposing the gel to UV light; detecting fluorescence emitted from the electrophoresis gel; excising at least one portion of the electrophoresis gel based upon the detected fluorescence, wherein said at least one portion contains proteins of the protein sample; and subjecting proteins from the at least one portion to mass spectroscopy. Using this method, more proteins can be identified by GeLC-MS than when the electrophoresis gel is treated with a protein stain or subjected to the gel handling steps accompanying such treatment.

    Abstract translation: 本文公开了一种制备用于质谱法的蛋白质样品的方法。 该方法包括在电泳凝胶上分离样品的蛋白质; 使蛋白质与卤素取代的有机化合物接触; 将凝胶暴露于紫外光下; 检测从电泳凝胶发出的荧光; 基于检测到的荧光来切割至少一部分电泳凝胶，其中所述至少一部分含有蛋白质样品的蛋白质; 并将至少一部分的蛋白质进行质谱分析。 使用这种方法，可以通过GeLC-MS鉴定更多的蛋白质，而不是用蛋白质染色剂处理电泳凝胶，或者进行伴随这种处理的凝胶处理步骤。

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