DIG LUMINESCENT DETECTION KIT

General description

DIG Luminescent Detection Kit is used for the detection of digoxigenin-labeled nucleic acids by enzyme immunoassay with luminescence on nylon membranes. The nonradioactive DIG system uses digoxigenin, a steroid hapten, coupled to deoxyuridine triphosphate (dUTP), UTP or didUTP to label DNA, RNA or oligonucleotides for hybridization and subsequent luminescent detection.
We are committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product is designed as a safer chemical. The DIG System was established as a sensitive and cost-effective alternative to using radioactivity for the labeling and detection of nucleic acids. There are many available publications that prove the versatility of the DIG System, so use of radio-labeling is no longer the only option for labeling of DNA for hybridization.

Application

DIG Luminescent Detection Kit has been used for highly sensitive and specific detection of DIG-labeled nucleic acids in:

• critical commercial assays
• Southern hybridization
• northern analysis
• plaque or colony lifts with anti-DIG-AP conjugate and the chemiluminescent substrate CSPD. Chemiluminescent detection with CSPD (following the instructions of the kit) is as sensitive as radioactive methods, but requires much shorter exposure times.

Packaging

1 kit containing 5 components.

Specifications

Sensitivity: 0.03 pg of homologous DNA or 0.1 pg of homologous RNA is detected in a Southern or dot blot after <30-min exposure time. Single-copy genes are detected in 0.3 μg mammalian genomic DNA.

Preparation Note

Working concentration: 0.5mg/ml for flow cytometry

• Storage conditions (working solution): Antibody conjugate (vial 3): once opened, should be stored at 2 to 8°C
• Blocking Reagent (bottle 4): in solution at 2 to 8°C
• CSPD (vial 5): at 2 to 8°C when frequently used. Repeated freeze/thaw cycles should be avoided.

Enzymatic dephosphorylation of the dioxetane CSPD by alkaline phosphatase leads to the metastable phenolate anion, which decomposes and emits light at 477nm. The light emission increases with time until a constant intensity is attained.
Assay Time

• Immunological detection 1.5 hours
• Signal detection 5 to 30 minute

Note: Store protected from light.

Other Notes

For life science research only. Not for use in diagnostic procedures.