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Beta-Galactosidase (β-Gal or X-Gal) staining kit

$375.00

Item Cat No.: BCBGAL

Application: Reporter assay

Reactivity: Tissue, Cell Culture

The β-Galactosidase staining kit determines the percentage of cells expressing lacZ reporter in cell cultures or frozen tissue sections.

1 Kit (100 slides)

LacZ is a bacterial gene that is often used as a reporter for the promoter activity of an eukaryotic gene. The lacZ gene can be transfected into cultured mammalian cells or inserted into the genome of transgenic animals by the CRISPR technology. The gene product of lacZ, β-galactosidase, catalyzes the hydrolysis of β-galactosides, i.e. X-gal, producing a blue color that can be visualized with bright-field microscopy.

BiCell Scientific’s β-Galactosidase staining kit provides a full set of reagents and protocols including fixation buffer and reaction buffer for staining β-Galactosidase in cell cultures or frozen tissue sections. Note that β-Galactosidase activities in FFPE specimens are difficult to detect because paraffin embedding at 60oC denatures β-Galactosidase proteins.

The kit contains:

  • 50ml of Fixation buffer (1.0% PFA, NaCl, and phosphate buffered saline) (Note that 1% PFA preserves β-Galactosidase activity while 3.7% PFA crosslinks proteins and reduces β-Galactosidase activity)
  • 50ml of Cryo-protection buffer (30% Sucrose and phosphate buffered saline)
  • 50ml of Staining buffer (K3[Fe(CN)6], K4[Fe(CN)6], MgCl2, and phosphate buffered saline)
  • 1ml of X-gal solution (50mg/ml in DMF)
  • 1ml of Mounting buffer (Mowiol and Tris buffered saline)

Store at -20oC


For Research Use Only. Not for use in clinical diagnostics.

  • Thaw kit content to room temperature.
  • For cultured cells, wash cells once with 1xPBS, add Fixation buffer and fix at 4oC for overnight.
  • For tissue sections, dissect out tissues and mince them into thin sagittal slices (2mm thick allowing easy fixative penetration). Drop-fix tissue slices in Fixation buffer at 4oC for overnight. Aspirate Fixation buffer and replace with Cryo-protection buffer. Incubate tissue slices at 4oC for overnight. Cut 5-10μm cryostat sections. Post-fix cryosections with Fixation buffer at 4oC for 10min right before staining experiment.
  • Wash fixed cells or tissue sections with 1xPBS twice.
  • Prepare β-Gal reaction solution by adding 10μl of X-gal solution to 500μl of Staining buffer.
  • Add β-Gal reaction solution to fixed cells or tissue sections.
  • Incubate samples at room temperature for overnight. (Note that reaction at room temperature has lower background than reaction at 37oC, because senescent cells have endogenous β-Gal activities.)
  • Drain off β-Gal reaction solution and wash stained cells or tissue sections with 1x PBS twice.
  • Add 20 ul of Mowiol solution to the section on each slide. Place a cover glass onto the section and apply a small pressure to remove any trapped air bubble.
  • Let Mowiol solution dry at room temperature for 1hr. Store glass slides at 4oC for less than 1 month.

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"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

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