Succinyl-CoA Synthetase Activity Assay Kit (Colorimetric) (ab196989)

Overview

  • Product name

    Succinyl-CoA Synthetase Activity Assay Kit (Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    Tissue, Adherent cells, Suspension cells, Purified mitochondria
  • Assay type

    Enzyme activity
  • Sensitivity

    < 100 µU
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Succinyl-CoA Synthetase Activity Assay Kit (Colorimetric) (ab196989) is an assay where Succinyl-CoA Synthetase (SCS) converts succinate into succinyl-CoA in the presence of ATP and CoA. Succinyl-CoA reacts with the SCS Developer to form a colored product with strong absorbance at OD=450 nm. This kit is simple to use and high throughput adaptable.

    It can detect less than 0.1 mU of Succinyl-CoA Synthetase activity in a variety of samples.

  • Notes

    Succinyl-CoA Synthetase (SCS, also called Succinyl-CoA ligase, Succinate Thiokinase) (EC 6.2.1.5) is a critical enzyme in the citric acid cycle (TCA) and an important metabolic intermediate for porphyrin, heme and ketone body biosynthesis. It is located in the mitochondrial matrix and is a heterodimer composed of one alpha and one beta subunit.

    In humans, Succinyl-CoA Synthetase deficiency causes the build-up of lactic acid leading to lactic acidosis, which can be fatal in infants. Measurement and analysis of SCS activity is useful for both mechanistic studies as well as for diagnostic purposes.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Developer 1 vial
    NADH Standard Yellow 1 vial
    Positive Control 1 vial
    SCS Assay Buffer 1 x 25ml
    SCS Enzyme Mix 1 vial
    SCS Substrate Mix 1 vial
  • Function

    Catalyzes the ATP-dependent ligation of succinate and CoA to form succinyl-CoA.
  • Tissue specificity

    Widely expressed. Not expressed in liver and lung.
  • Pathway

    Carbohydrate metabolism; tricarboxylic acid cycle; succinate from succinyl-CoA (ligase route): step 1/1.
  • Involvement in disease

    Mitochondrial DNA depletion syndrome 5
  • Sequence similarities

    Belongs to the succinate/malate CoA ligase beta subunit family.
    Contains 1 ATP-grasp domain.
  • Cellular localization

    Mitochondrion.
  • Information by UniProt
  • Alternative names

    • ATP-specific succinyl-CoA synthetase subunit beta
    • mitochondrial
    • Renal carcinoma antigen NY-REN-39
    • SCS-betaA
    • SUCB1_HUMAN
    • Succinyl-CoA ligase [ADP-forming] subunit beta
    • Succinyl-CoA synthetase beta-A chain
    • SUCLA2
    see all

Images

  • Typical NADH Standard Curve using colorimetric reading.

  • Succinyl-CoA Synthetase activity measured in mitochondria isolated from bovine heart, yeast (P. pastoris) and MCF-7 cells and in Jurkat cell lysate. The graph also shows data from the SCS Positive Control included in the kit and Blank background baseline.

  • Succinyl-CoA Synthetase specific activity in mitochondria prepared from yeast (P. pastoris, 1.14 μg), bovine heart (1.1 μg) and MCF-7cells (0.5 μg), and in Jurkat cell lysate (25 μg).

Protocols

References

ab196989 has not yet been referenced specifically in any publications.

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