beta Galactosidase IHC Detection Kit

Product Discontinued

beta Galactosidase IHC Detection Kit Summary

• Description: The B-galactosidase staining kit contains the necessary reagents to perform 100 assays in a 6-well plate format, providing an easy and rapid method to determine transfection efficiency by indicating beta -Gal activity in individual intact cells. The transfected cells can be stained in tissue culture dishes without having to remove or manipulate the cells.
• Kit Type: IHC Detection Kit

Applications/Dilutions

• Dilutions:
  • Cellular Imaging
  • Enzyme Activity
  • Immunohistochemistry

Packaging, Storage & Formulations

• Storage: Storage is content dependent.

Notes

Additional items required (not included in the kit):
6-Well Tissue Culture Plates (Or tissue culture plates of choice)
Cells transfected with an expression plasmid encoding the LacZ gene for B-Gal expression.
Untransfected Cells for a background control.
Pipettor
37C incubator
Light Microscope
1X PBS (approximately 10 mL per well for the 6-well format)

Alternate Names for beta Galactosidase IHC Detection Kit

• EC 3.2.1.23; b-gal

Background

The LacZ gene encodes the Beta-Galactosidase (Beta-Gal) enzyme that catalyzes the hydrolysis of B-galactosides, including lactose. The LacZ gene is among the most widely used and published reporter genes due in large part to the extreme resistance of the B-Gal enzyme to proteolytic degradation. Beta-Gal enzyme activity is also readily detected in stably or transiently transfected cells or transgenic tissues using a variety of substrates. The beta-Gal Staining Kit is based on 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal), a noninducing chromogenic substrate for beta-Gal. When beta-Gal cleaves the glycosidic linkage in X-Gal, a soluble, colorless indoxyl monomer is produced. Subsequently, two of the liberated indoxyl moieties form a dimer which is nonenzymatically oxidized. The dimerizatation and oxidation reactions require transfer of an electron which is facilited, in this case, by ferric and ferrous ions. The resultant halogenated indigo is a very stable and insoluble dark blue compound. Following transfection with plasmids that contain LacZ, and the addition of X-Gal, the transfection efficiency can be determined by counting the number of blue cells in the total cell population.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for beta Galactosidase Kit (NBP2-29546)(1)

Publication using NBP2-29546

• Alpert E, Gruzman A, Tennenbaum T et al. Selective cyclooxygenase-2 inhibitors stimulate glucose transport in L6 myotubes in a protein kinase Cdelta-dependent manner. Biochem Pharmacol. 2007-02-01 [PMID: 17098211]
  
  Details:
  On fixed cells.