Hitrap 5 ml desalting columns

Example 1

Solutions of stannous chloride (40 mg/mL, Sigma 243523) were prepared in deionized water under flowing nitrogen. A 0.5 mL aliquot was filtered and mixed with 1.00 mL NaCl (1.00 M) plus 150 mg of preservative-free heparin (10,000 IU/mL). Approximately 100 mCi of freshly eluted ^99mTc was added and mixed for 30 minutes at room temperature. Aliquots containing approximately 10 mCi of ^99mTc and 20 mg of heparin were removed for tissue experiments. FIG. 3A shows the labeling affinity, measured by paper chromatography Whatman number 35 with acetone, showed more than 97% binding of heparin to ^99mTc.

Radiolabeled heparin was also analyzed by Sephadex G25 column chromatography (HiTrap 5 mL desalting columns, GE healthcare, 17140801) with 0.15 M NaCl as the elution buffer, and approximately 1 mL fractions were collected. FIG. 3B shows radioactivity eluted at fraction 4 (free ^99mTc elutes at fraction 13), demonstrating that all of the ^99mTc eluted at the void volume and confirming that there is no unbound ^99mTc in the radiolabeled heparin. The stability of ^99mTc-heparin in an acidic environment was tested by diluting in artificial gastric juice (Carolina, 865603) and showing that is properties were unaltered, using both paper chromatography and Sephadex G25.

Example 1

Solutions of stannous chloride (40 mg/mL, Sigma 243523) were prepared in deionized water under flowing nitrogen. A 0.5 mL aliquot was filtered and mixed with 1.00 mL NaCl (1.00 M) plus 150 mg of preservative-free heparin (10,000 IU/mL). Approximately 100 mCi of freshly eluted ^99mTc was added and mixed for 30 minutes at room temperature. Aliquots containing approximately 10 mCi of ^99mTc and 20 mg of heparin were removed for tissue experiments. FIG. 3A shows the labeling affinity, measured by paper chromatography Whatman number 31 with acetone, showed more than 97% binding of heparin to ^99mTc.

Radiolabeled heparin was also analyzed by Sephadex G25 column chromatography (HiTrap 5 mL desalting columns, GE healthcare, 17140801) with 0.15 M NaCl as the elution buffer, and approximately 1 mL fractions were collected. FIG. 3B shows radioactivity eluted at fraction 4 (free ^99mTc elutes at fraction 13), demonstrating that all of the ^99mTc eluted at the void volume and confirming that there is no unbound ^99mTc in the radiolabeled heparin. The stability of ^99mTc-heparin in an acidic environment was tested by diluting in artificial gastric juice (Carolina, 864603) and showing that its properties were unaltered, using both paper chromatography and Sephadex G25.

Example 5

Solutions of stannous chloride (40 mg/mL, Sigma 243523) were prepared in deionized water under flowing nitrogen. A 0.5 mL aliquot was filtered and mixed with 1.00 mL NaCl (1.00 M) plus 150 mg of preservative-free heparin (10,000 IU/mL). Approximately 100 mCi of freshly eluted ^99mTc was added and mixed for 30 minutes at room temperature. Aliquots containing approximately 10 mCi of ^99mTc and 20 mg of heparin were removed for tissue experiments.

Results. Labeling affinity was measured by paper chromatography Whatman number 31 with acetone, which confirmed greater than 97% binding of heparin to ^99mTc.

The heparin was also analyzed by Sephadex G25 column chromatography (HiTrap 5 mL desalting columns, GE healthcare, 17140801) with 0.15 M NaCl as the elution buffer, and approximately 1 mL fractions were collected. The test demonstrated that all of the ^99mTc eluted at the void volume and confirmed that there was no unbound ^99mTc in the radiolabeled heparin.

The stability of ^99mTc-heparin in an acidic environment was tested by diluting in artificial gastric juice (Carolina, 864603) and showing that its properties were unaltered, using both paper chromatography and Sephadex G25.