Overview

  • Product name

    FACS Blue LacZ beta Galactosidase detection kit
    See all beta galactosidase kits
  • Product overview

    Although chromogenic assays of β-galactosidase activity (i.e. X-Gal) have use, application of the fluorogenic substrate 3-carboxyumbelliferyI β-D-galactopyranoside (CUG) (blue fluorescence) combined with Fluorescence Activated Cell Sorting (FACS) analysis has been shown to be several orders of magnitude more sensitive. In addition, because of its high water solubility and detection limits, the CUG substrate has found extensive use in automated ELISA type assay systems. Abcam’s FACS Blue LacZ beta Galactosidase detection kit (ab189815) is especially useful for dual-labeling experiments where the cells to be analyzed are also labeled with a fluorescein based probe (FITC-labeled antibody, fluorescein-based substrate, etc.).

    Emission: 460 nm.

    Excitation: 390 nm.

  • Notes

    One of the most common reporter genes used in molecular biology applications is the E. coli lacZ gene that codes for an active subunit of β-galactosidase in vivo. Since this enzyme is generally absent in normal mammalian, yeast, some bacterial and even plant cells, it can be detected at very low levels, and since the enzyme has a wide substrate specificity, monitoring lacZ expression (and therefore co-expressed genes or promoter efficiency) has become routine to the point of detection of as few as 5 copies of β-galactosidase per cell by FACS analysis.

    Abcam’s FACS Blue LacZ beta Galactosidase detection kit (ab189815) provides reagents and protocol to perform up to 500 tests (10 x 96 well microtitre plates).

  • Platform

    Flow cytometer

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 500 tests
    Inhibitor 1 x 1ml
    Reference Standard 1 x 0.5ml
    Substrate Reagent 1 x 0.5ml
  • Research areas

  • Relevance

    Beta galactosidase is a hydrolase enzyme that cleaves beta-linked terminal galactosyl residues from gangliosides, glycoproteins, and glycosaminoglycans. Beta galactosidase is an essential enzyme in the human body. Deficiencies in the protein can result in galactosialidosis or Morquio B syndrome. Senescent cells display senescence-associated expression of beta galactosidase activity.
  • Cellular localization

    Isoform 1: Lysosome. Isoform 2: Cytoplasm, perinuclear region. Note=Localized to the perinuclear area of the cytoplasm but not to lysosomes.
  • Alternative names

    • Acid beta galactosidase
    • Beta galactosidase 1
    • EBP
    • ELNR1
    • Galactosidase beta 1
    • GLB1
    • MPS4B
    see all

Protocols

References

This product has been referenced in:

  • Timraz SB  et al. In-depth evaluation of commercially available human vascular smooth muscle cells phenotype: Implications for vascular tissue engineering. Exp Cell Res 343:168-76 (2016). Read more (PubMed: 27079869) »
See 1 Publication for this product

Customer reviews and Q&As

Answer



In regards to your question on reagentand preparation, the substrate reagent issupped at 50 mM (and comes as distilled water, for your information only no adding of water is necessary), you add 0.5 ml of the reagent to 2 ml of reaction buffer to create the 10 uM Substreate mix you need to use in the assay.



The reference standard is supplied as 20 uM (and comes in methanol already), and you dilute with stop reagent to optimize the spectometer conditions, a 1:50 dilution is typically enough.

The inhibitor is supplied as 30 mM in distilled water and is used as need in the protocol instructions.

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