ZytoLight^® SPEC PTEN/CEN 10 Dual Color Probe

The ZytoLight^® SPEC PTEN/CEN 10 Dual Color Probe (PL37) is a direct-labeled fluorescence in situ hybridization (FISH) probe set containing green and orange labeled polynucleotides targeting sequences in the PTEN gene region and chromosome 10 alpha satellite centromeric region in formalin-fixed, paraffin-embedded (FFPE) tissue specimens. This probe utilizes FISH technology for the visualization of specific nucleic acid sequences.

Principle:

Fluorescence in situ hybridization (FISH) detects and visualizes specific nucleic acid sequences in cell preparations. Fluorescently-labeled DNA fragments (FISH probes) co-denature and hybridize with complementary target DNA strands, followed by stringent washing to remove unbound probes. The hybridized probes are visualized using a fluorescence microscope with specific excitation and emission filters.FISH assesses the copy number of PTEN and CEN10 using direct probes that hybridize after denaturation, with unbound probes washed away and a DAPI counterstain applied. The signals—green for PTEN and orange for CEN10—indicate deletions. This antibody-free method is optimized for fluorochromes, aiding in the identification of tumor suppressor gene aberrations.

Components:

ZytoLight^® SPEC PTEN/CEN 10 Dual Color Probe comprises:

• ZyGreen labeled polynucleotides (excitation 503 nm/emission 528 nm) targeting the PTEN gene region on chromosome 10 (10q23.2-q23.31).
• ZyOrange labeled polynucleotides (excitation 547 nm/emission 572 nm) targeting the alpha satellite centromeric region D10Z1 of chromosome 10.k
• Formamide-based hybridization buffer.

The probe is designed for qualitative detection of sequences in the PTEN gene region and chromosome 10 centromere in formalin-fixed, paraffin-embedded (FFPE) tissue specimens by FISH. It is used in combination with the ZytoLight^® FISH-Tissue Implementation Kit for specimen pretreatment, hybridization, and post-hybridization processing.

For the qualitative detection of human PTEN gene deletions and alpha satellites of chromosome 10 by fluorescence in situ hybridization (FISH)

In a normal interphase nucleus, the expected result is the presence of two orange and two green signals. If there is a deletion of the PTEN gene locus, a reduced number of green signals will be observed. Variations in signal patterns may indicate different genetic aberrations and should be investigated further. Proper specimen handling and processing are crucial to avoid artifacts or false results.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany