Clarity OTX Lysis-Loading Buffer v2.0, 1L, Clarity OTX Lysis-Loading Buffer v2.0, 1L, Ea

Бренд: Phenomenex

General Extraction

Common Extraction Procedure (~180 minutes)

Suitable for Most Oligo Therapeutics & Samples
Oligo types Sample Types
DNA Plasma
Aptamers Serum
RNAi/siRNA Urine
Thioates Tears
Lipid-conjugates Saliva
Liposome encapsulated Tissue

Extraction Protocol

Sample Pre-Treatment
Tissue
  • Proteinase K digestion or homogenize
  • Mix 1-to-1 with Lysis-Loading buffer
Biological Fluids
  • Mix 1-to-1 with Lysis-Loading buffer

Detect Low Dosage Levels

Due to the typical 80 % and greater recoveries of the parent oligonucleotide therapeutic and its metabolites, detection in the picomole range is possible. The TIC below illustrates detection of a dosage in this very sensitive range. Aliquots of 500 ng & 50 ng spiked into plasma were extracted using Clarity OTX to determine sensitivity limits. A TIC of the 500 ng load is shown below. The 14.3 minute peak corresponding to the 19mer P-S can be quantitated. While a peak is not observed for the TIC of the 50 ng load, using a XIC at m/z of 944 (the -7 charge state) one can still quantitate the oligonucleotide at very low levels.

Sensitivity Study in Plasma

Column:
Clarity 3 µm Oligo-RP C18
Dimension:
50 x 4.6 mm
Part No.:
00B-4441-E0
Mobile Phase:
A: 50 mM TEAA, pH 7.5 / 5 % Acetonitrile B: Methanol
Gradient:
A/B (90:10) to A/B (40:60) in 20 min
Flow Rate:
1 mL/ min
Detection:
UV @ 260 nm
Sample:
25nt DNA oligonucleotide
UV chromatograms of oligonucleotide extracted from liver tissue using Clarity OTX. The 19mer extracted phosphorothioate oligonucleotide was spiked with 10 µg of a oligonucleotide internal standard before analysis. The top two chromatograms represent different levels of the incubated P-S oligo. The bottom chromatogram is an external standard of equal amounts of the 19mer oligo and internal standard. Note the high recovery of the oligonucleotide and low level of plasma contaminants from the incubated samples.

Sensitivity Study in Live Tissue

Артикул: AL0-8579 Категория: Бренд: