Overview

  • Product name

    Glycine Assay Kit (Fluorometric)
  • Detection method

    Fluorescent
  • Sample type

    Saliva, Urine, Serum, Plasma, Cell Lysate, Tissue Lysate
  • Assay type

    Quantitative
  • Sensitivity

    1 µM
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Glycine Assay Kit (Fluorometric) (ab211100) provides a provides a simple, sensitive, and high-throughput adaptable assay that detects physiological concentrations of glycine (GLY) in multiple biological samples such as cell and tissue lysates and biological fluids. In this assay, Glycine is oxidized in the presence of GLY Enzyme Mix is converted to an intermediate, which reacts with the fluorescent probe to generate a strong stable signal at Ex/Em = 535/587 nm. The intensity of the signal is directly proportional to the amount of GLY in the sample.


    The reaction is specific and other amino acids do not interfere with the assay. The assay can detect as little as 1 µM of Glycine in a variety of biological samples.

  • Notes

    Glycine (GLY, G) is one of the 20 standard amino acids commonly found in proteins. Glycine’s side chain is a hydrogen substituent, which makes it the smallest proteogenic and the only non-chiral amino acid. Basic functions of Glycine include the participation in the synthesis of creatine, glutathione, heme groups, and conjugated bile acids (bile salts). It is also present as one of the most abundant residues in the triple-helical structure of collagen.

    Glycine acts as a glucogenic amino acid by regulating sugar levels in blood. Therefore, glycine supplementation has been used in patients suffering anemia, hypoglycemia and chronic fatigue. Glycine possesses both inhibitory and excitatory neurotransmitter functions in the brain stem and spinal cord. In cancer cells, glycine consumption is highly correlated to cancer cell proliferation via purine synthesis. Glycine uptake in cancer cell studies supports the role of this amino acid in tumorigenesis and malignancy.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    GLY Assay Buffer 1 x 25ml
    GLY Developer (200 µg) 1 vial
    GLY Enzyme Mix (600 mU) 1 vial
    GLY Probe 1 x 400µl
    GLY Standard (10 µM) 1 vial
  • Research areas

  • Relevance

    Defects in GLDC are a cause of nonketotic hyperglycinemia (NKH), also known as glycine encephalopathy (GCE). NKH is an autosomal recessive disease characterized by accumulation of a large amount of glycine in body fluid and by severe neurological symptoms. The degredation of glycine is catalised by the glycine cleavage system. The P protein binds the alpha-amino group of glycine through its pyridoxal phosphate cofactor; carbondioxide is released and the remaining methylamine moiety is then transferred to the lipoamide cofactor of the H protein. The glycine cleavage system is composed of four proteins: P, T, L and H.

Images

  • Typical Glycine standard calibration curve.

  • Estimation of Glycine in human serum, saliva, and urine. Samples were deproteinized using 10 kD spin column (ab93349) and diluted with GLY Assay Buffer prior to performing assay: (serum 1:64 dilution; saliva 1:32 dilution; urine 1:128 dilution). 25 µL of each diluted sample was spiked with 0.3 nmol of Glycine Standard and assayed following the kit protocol. Glycine concentrations in serum (224 ± 21µM), saliva (149 ± 7 µM) and urine (54 ± 4 µM/mM creatinine).

Protocols

References

ab211100 has not yet been referenced specifically in any publications.

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