ZytoLight^® SPEC VHL/1p12/CEN 7/17 Quadruple Color Probe

The ZytoLight SPEC VHL/1p12/CEN 7/17 Quadruple Color Probe (PL60) is a direct-labeled FISH probe designed for the qualitative detection of deletions involving the human VHL gene, chromosome 1p12 specific sequences, and alpha satellites on chromosomes 7 and 17 in formalin-fixed, paraffin-embedded (FFPE) specimens. It is intended for use with the ZytoLight FISH-Tissue Implementation Kit.

Principle:

The fluorescence in situ hybridization (FISH) technique allows for the detection and visualization of specific nucleic acid sequences. Fluorescently-labeled DNA fragments (FISH probes) co-denature and hybridize with complementary target DNA strands. After stringent washing to eliminate unbound fragments, the hybridized probes are visualized using a fluorescence microscope with specific excitation and emission filters.Fluorescently labeled locus-specific and centromeric DNA probes co-denature with target DNA in pretreated formalin-fixed, paraffin-embedded (FFPE) sections. They hybridize overnight after denaturation at 75°C, followed by washing and DAPI counterstaining, then are visualized with color-specific filters.For the assay, apply 10 µl of the probe to each slide, seal with a coverslip and rubber cement, denature for 10 minutes at 75°C, hybridize at 37°C, and process according to ZytoLight FISH-Tissue Implementation Kit instructions.

Components:

ZytoLight^® SPEC VHL/1p12/CEN 7/17 Quadruple Color Probe comprises:

• ZyGreen labeled polynucleotides (excitation 503 nm/emission 528 nm) targeting the VHL gene region on chromosome 3 (3p25.3).
• ZyRed labeled polynucleotides (excitation 580 nm/emission 599 nm) targeting chromosome 1p12.
• ZyGold labeled polynucleotides (excitation 532 nm/emission 553 nm) targeting chromosome 7.
• ZyBlue labeled polynucleotides (excitation 418 nm/emission 467 nm) targeting chromosome 17.
• Formamide-based hybridization buffer.

For the qualitative detection of the human VHL gene, chromosome 1p12 specific sequences, and alpha satellites of chromosomes 7 and 17 by fluorescence in situ hybridization (FISH)

The probe set enables qualitative detection of the VHL gene region, 1p12 locus sequences, and centromeres of chromosomes 7 and 17 in formalin-fixed, paraffin-embedded (FFPE) tissue specimens by FISH. Used with ZytoLight^® FISH-Tissue Implementation Kit for specimen pretreatment, denaturation/hybridization, and post-hybridization processing.

In a normal interphase nucleus, two green signals (for VHL), two red signals (for chromosome 1p12), two gold signals (for chromosome 7), and two blue signals (for chromosome 17) are expected. In cases of deletion or amplification, abnormal signal patterns may be observed, which should be investigated further.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany