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FLOWSCRIPT® HPV E6/E7 assay kit

Flow cytometry-based assay for the single cell detection of mRNAs for HPV oncoproteins E6 and E7
 
ENZ-GEN300-0100 100 tests Inquire for pricing
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  • Homogeneous system eliminates post-hybridization wash steps, minimizing leakage and signal degradation
  • Routine use of external controls enables consistent assay performance
  • Compatible with high throughput testing platforms
  • Low input volume saves precious samples for additional analyses
The FLOWSCRIPT® HPV E6/E7 assay kit is a flow cytometry based assay for the detection of E6/E7 mRNA in whole cells. The assay employs a novel in situ hybridization technique utilizing cocktails of oligonucleotide probes specific to multiple targets within the E6 and E7 genes to ensure the detection of these transcripts from most known variants of high risk HPV. Fixed and permeabilized cells are hybridized with probes and analyzed by flow cytometry for E6/E7 transcript expression. Each probe contains a fluorescent label and a quenching molecule whereby no signal is observed in the absence of target.  During hybridization, the probe anneals to the target sequence, thereby emitting a detectable signal.
ENZ-GEN300 schematic
The human HPV genome contains 8 open reading frames which include E6, E7, E1, E2, E4, E5, L2 and L1. "E" or "L" refer to "early" or "late" functions, respectively. During the course of cancer development, the viral molecule frequently becomes integrated into host-cell DNA.
ENZ-GEN300 workflow
FLOWSCRIPT™ HPV E6/E7 Assay Workflow
ENZ-GEN300 flow
Typical expected results. Negative control cells (ENZ-GEN302) and Positive control cells (ENZ-GEN301) were assayed using the FLOWSCRIPT™ E6/E7 HPV Assay. Negative control cells (left panel) that over-express oncoprotein E6/E6 mRNA comprise < 5% of cells in the analysis gate. Positive control cells that over-express oncoprotein E6/E7 mRNA comprise > 95% of cells in the analysis gate.
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ENZ-GEN300 schematic ENZ-GEN300 workflow ENZ-GEN300 flow

Product Specification

Assay Time:<3 hours
 
Applications:Flow Cytometry, Fluorescent detection
 
Application Notes:The FLOWSCRIPT® HPV E6/E7 Assay is compatible with any properly calibrated cytometer with a laser and detector setup capable of exciting and reading emissions from fluorescein. It has been validated for use on the following instruments: BD Accuri™, BD FACSCalibur™, BD FACSVerse™, guava easyCyte™, handyem™ HPC150, and ACEA NovoCyte™.
 
Specificity:The FLOWSCRIPT® HPV E6/E7 Assay is capable of detecting E6/E7 mRNA transcripts from multiple high risk HPV types: HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 82.  Together, these types account for over 95% of cervical cancer.
 
Quantity:100 flow cytometry assays
 
Use/Stability:FLOWSCRIPT® probe cocktails are stable for 6 months at -20°C.  Buffers are stable for 6 months at 2-8°C.
 
Shipping:Shipped on Blue Ice
 
Kit/Set Contains:Buffer A (Fixation Buffer)
Buffer B (Hybridization Buffer)
HPV E6/E7 Probe Cocktail
 
Scientific Background:
 
Technical Info/Product Notes:For FLOWSCRIPT® HPV E6/E7 positive control cells, see Prod. No. ENZ-GEN301.
For FLOWSCRIPT® HPV E6/E7 negative control cells, see Prod. No. ENZ-GEN302.

Application Notes:
HPV E6/E7 mRNA Expression Analysis Using the NovoCyte® Flow Cytometer

FLOWSCRIPT® Platform Detects Genotypic and Phenotypic Markers Associated with HPV Infection and Cancer Progression
 

General Literature References

Human papillomavirus testing in the prevention of cervical cancer: M. Schiffman, et al.; J. Natl. Cancer Inst. 103, 368 (2011), Abstract; Full Text
Human papillomavirus and cervical cancer: M. Schiffman, et al.; Lancet 370, 890 (2007), Abstract;

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