echemi logo
Product
  • Product
  • Supplier
  • Inquiry
    Home > Biochemistry News > Biotechnology News > Trans-amino action (paper lithography)

    Trans-amino action (paper lithography)

    • Last Update: 2020-10-30
    • Source: Internet
    • Author: User
    Search more information of high quality chemicals, good prices and reliable suppliers, visit www.echemi.com

    one. Objectiveto understand
    the
    of trans-aminos in animal-based tissues and to learn the paper-based analysis identification method of trans-amino reaction products.two. Principle
    the main way
    breakdown of metabolism in the body is deamination. The main ways of deamination are oxidizing deamin, trans-amino, combined deaminizing and non-oxidizing deamination. Trans-amino refers to the interchange reaction between α-amino acids and α ketones of α-ketoic acids, catalyzed by
    transaminase
    . Transaminase is widely distributed in various organ tissues of animal bodies, and all kinds of transaminases have a single transaminase catalysis.experiment, glutamate and acetone were estrogenated with liver plasma to produce alanine under the catalyst of alanine transaminase in liver cells. The presence of alanine was identified by paper-based analysis to prove the transamination of tissues. In order to prevent the substrate acetone acid from being oxidized or reduced by other enzymes in the tissue, iodized acetic acid (or bromoacetic acid) inhibits the enzyme and oxidation of sugar.


    .


    .


    .
    results: α and ketone diacine did not react
    paper-based analysis was used to determine the presence or absence of trans-aminoline product alanine. The principle is: glutamate and alanine are two amino acids with different chemical properties, the former is hydrophobic amino acids, the latter is hydrophobic amino acids, according to which the two in the fixed phase (water) and flow phase (phenol
    reagents
    ) in different degrees of distribution, so the flow rate is different. Finally,three ketones to color.
    wednesday. Materials
    (i) instruments
    test tubes
    ;
    centrifuges
    ;
    centrifuges
    ; scissors and tweezers;
    culture
    dishes; surface dishes; capillary tubes; sprayers; hair dryers.
    (ii) Reagent
    1. 0.1% Alanine solution
    2. 1% glutamate solution
    3. 1% sodium acetone solution
    4. 0.25% iodine acetic acid solution
    5. 0.01 mol/LpH7.4 phosphoric acid buffer
    6. 0.1%tritone ethanol solution
    7. The layering solvent (the saturated water solution of phenol)
    four. Methods
    (i) Preparation of liver homogenous slurry to take 1 g of fresh animal liver, cut in a research, with a little fine sand, 5 ml pre-cooled 0.01 mol/LpH7.4 phosphoric acid buffer, ground into a homogenous slurry.
    (ii) Trans-amino reaction
    1. Take 2 small test tubes, respectively marked as assay and control, each plus liver plasma 10 drops, the control tube immediately put in a boiling water bath for 10 minutes, remove cooling.
    2. Two tubes each plus 1% glutamate 10 drops, 1% sodium acetone 10 drops, 0.25% iodine acetic acid 5 drops, put 40 degrees C water bath insulation after 30 minutes to remove.
    3. Two tubes each plus 5% triclosal acetic acid 2 drops, in a boiling water bath for 5 minutes (2000 rpm), the upper liquid will be moved into the same number of small test tube spare.
    (iii) Layering verification
    1. Take a 13cm diameter circular filter paper, with a pencil over the center of the circle for two vertical lines, in each line from the center of the circle 1cm to mark, as a sample, and at its edge marked, valley, pair, C words, see Figure 9.
    2. The measuring solution, 1% glutamate solution, control solution and 0.1% alanine solution were taken with capillary glass tubes respectively, and the corresponding sample on the filter paper was sampled at 2 μl, with a diameter of no more than 3mm.
    3. With a large needle in the center of the filter paper round hole (about 1mm diameter), take another filter paper (0.5×2.5cm), rolled into a barrel- shape, tight as a wick, from the filter paper on the corresponding point sample back insertion hole, highlighting the filter paper about 1cm.
    4. Add about 2.5 ml of laminate solvent in a surface dish with a diameter of 5 cm, the surface dish is placed in the middle of a petri dish with a diameter of 10cm, the filter paper is flat on the petri dish, so that the filter paper element immersed in the layer solvent, take another dish of the same size reverse cover. The laminate solvent rises along the filter paper element to the filter paper and spreads around it, waiting for the solvent front to be removed 1.5cm from the edge of the filter paper (about 50 minutes for the paper). Dry with a hairdryer or electric stove after drawing the solvent front with a pencil.
    5. ColorIng Place the filter paper flat on a Petri dish, spray 0.1%tritone ethanol solution, and dry it with a hairdryer or electric oven. Observe the curved purple spots that appear in the layering and draw them out with a pencil. Calculates the RF value of the various color spots.
    : Before touching the filter paper in operation, wash your hands and wipe them clean, and try to avoid contact between your hands and the middle part of the filter paper. .
    This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only. This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed description of the concern or complaint, to service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content will be removed immediately.

    Contact Us

    The source of this page with content of products and services is from Internet, which doesn't represent ECHEMI's opinion. If you have any queries, please write to service@echemi.com. It will be replied within 5 days.

    Moreover, if you find any instances of plagiarism from the page, please send email to service@echemi.com with relevant evidence.