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    Home > Biochemistry News > Biotechnology News > Gel filtration separates proteins.

    Gel filtration separates proteins.

    • Last Update: 2020-10-30
    • Source: Internet
    • Author: User
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    related topics .Objectives:
    1, learning the principles and operating methods of gel
    column
    technology;
    2, learning hemoglobin preparation methods.II, experimental principle:
    1. Gel layering: principle and application
    2. Principle of reaction to this experiment: Hemoglobin (hemoglobin, Hb) is the main inclusion of red blood cells, it is hemoglobin and globin peptide chains connected by a binding protein, is a pigment protein. Hemoglobin (Mw-64 500) is added to excess potassium cyanide (K3Fe(CN) 6, Mw-327.25), and hemoglobin reacts with high-iron potassium cyanide to produce high iron hemoglobin (MetHb). In order to remove excess high-speed iron potassium cyanide, get a purer MetHb sample, the above mixture through the cross-linked glucosaccharide gel column, with phosphoric acid buffer washed away, from different colors, can be directly observed MetHb (reddish-brown) washed away faster, and
    small molecules
    high-iron potassium cyanide (yellow) washed out slowly, to achieve the goal of completely separating MetHb.II, experimental materials,
    reagents
    and instruments:
    (i) materials: chicken anticoagulant whole blood
    (ii) reagents:
    1. Carbon tetrachloride (CCl4)
    2. Physiological saline
    3. Cross-linked glucosamine G-25 (fine grain)
    4. 0.1mol/L
    phosphate buffer
    (pH7.0):
    a, 0.1M sodium dihydrophosphate; b, 0.1M sodium phosphate dihydrophosphate;
    a, b mixed at a ratio of 49:51, and proofed
    5. 0.4% K3Fe (CN)6.
    6. Cold physiological saline
    (iii) instrument (slightly)





    , experimental steps:
    1. Treatment of the gel:
    take 4 grams of Sephadex-G25 dry powder, soak in distilled water fully soothing (room temperature, 6 hours), then tilt the method to remove the surface suspension of small particles, and finally add the same volume pH7 phosphoric acid buffer to continue soaking.
    .
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