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  • Bradford Assay For Protein Quantification For WB First Make Standards

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    alrains
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    The document provides instructions for performing a Bradford assay to quantify protein concentrations for western blotting. It includes making a standard curve with BSA concentrations ranging from 0-2000 μg/mL. Samples are diluted 5-fold before adding 5 μL to wells containing Bradford dye. Absorbance is then read at 595 nm and sample concentrations can be determined based on the standard curve.

    Исходное описание:

    Quantification of proteins

    Оригинальное название

    BFA Protein Quant

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    The document provides instructions for performing a Bradford assay to quantify protein concentrations for western blotting. It includes making a standard curve with BSA concentrations ranging from 0-2000 μg/mL. Samples are diluted 5-fold before adding 5 μL to wells containing Bradford dye. Absorbance is then read at 595 nm and sample concentrations can be determined based on the standard curve.

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    © All Rights Reserved
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    2 просмотров1 страница

    Bradford Assay For Protein Quantification For WB First Make Standards

    Загружено:

    alrains
    The document provides instructions for performing a Bradford assay to quantify protein concentrations for western blotting. It includes making a standard curve with BSA concentrations ranging from 0-2000 μg/mL. Samples are diluted 5-fold before adding 5 μL to wells containing Bradford dye. Absorbance is then read at 595 nm and sample concentrations can be determined based on the standard curve.

    Авторское право:

    © All Rights Reserved
    Скачайте в формате DOCX, PDF, TXT или читайте онлайн в Scribd
    Отметить как неприемлемый контент
    из 1

    BRADFORD ASSAY FOR PROTEIN QUANTIFICATION FOR WB

    First make standards


    Tube#
    1
    2
    3
    4
    5
    6
    7
    8 (blank)

    Standard
    Volume (L)
    10
    30
    20
    20
    20
    20
    20
    -

    Source of
    Standard
    2mg/ml stock
    0
    2mg/ml stock
    10
    2mg/ml stock
    20
    Tube 2
    20
    Tube 3
    20
    Tube 5
    20
    Tube 6
    20
    20

    Diluent
    Final
    Solume (L)
    [Protein](g/mL)
    2000
    1500
    1000
    750
    500
    250
    125
    0

    Steps
    1.
    2.
    3.
    4.
    5.
    6.
    7.

    Mark wells (standards/samples either in triplicates or duplicates)


    Prepare standards as per the table given above
    Dilute samples x5 times ie 2 L of sample + 8 L of ddH2O
    Add 250 L of Bradford dye to all wells
    Add 5 L of standards / samples to the designated wells
    Mix well and remove all the bubbles
    Read at absorbance 595 mm

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