Actin RNA Probe, DIG-labeled

This antisense RNA probe (human β-actin) is in vitro transcribed in the presence of digoxigenin (DIG)-UTP. The transcript has a length of 588 bases. 550 bases are complementary to the 5′ region of human β-actin mRNA (nucleotides 69 to 6l8, EMBL: HSAC07). The additional 25 bases at the 5′ end, and the 13 additional bases at the 3′ end of the transcript are specific for the promoter/polylinker region of the transcription vector.

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 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.

Actin RNA Probe, DIG-labeled, is specifically useful for evaluating the quality and quantity of various RNA species and can be used:

• in In situ hybridization (for example, as a control in mRNA detection)
• for quality control in the construction of cDNA libraries
• in northern blot analysis to evaluate RNA from various human cell lines and tissue samples

Actin RNA Probe, DIG-labeled, is supplied in autoclaved, DEPC-treated water.

Agarose gel electrophoresis under denaturing conditions and subsequent northern blot analysis reveal a defined band of 588 bases.

Working solution: Dilute Sample

• Remove the desired number of isotyping strips from the canister. Remove the caps from an equal number of development tubes.
  The tubes may be labeled with a pencil or felt-tipped lab marker for easy identification.
• Dilute a sample containing the mouse monoclonal antibody in 1% BSA/ phosphate-buffered saline (PBS), pH 7.2 to 7.6.
  Culture supernatant samples should be diluted 1:10 to 1:100. Ascites samples should be diluted 1:20,000. These are recommended dilutions and may vary depending on the concentration of antibody in your sample. In our experience, a monoclonal antibody
  concentration of 0.1 to 1 g/ml of diluted sample gives the best results. 150 ml of this diluted sample will be added to the development tube.

Storage conditions (working solution): During assay, the preparation should be maintained at 0 °C.

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