Labeling efficiency was assessed by instant thin layer chromatography (iTLC) using silica gel coated paper (Varian Medical Systems, Inc.) and 0.1 M citrate buffer pH 5 as eluent. Colloid formation was determined by iTLC using silica gel-coated paper and 1 M NH4OAc:methanol (1:3) as eluent. Radiochemical purity of labeled peptides was analyzed by RP-HPLC on a Breeze system (Waters). A C-18 column (Symmetry Shield, 4.6 mm x 250 mm; particle size 5 μm, Waters) was used at a flow rate of 1 mL/min with the following buffer system: buffer A, 0.1% v/v trifluoroacetic acid in water; buffer B, methanol; with a gradient as follows: 100% buffer A (0-5 min), 60% buffer B (5-5.01 min), 80% buffer B (5.01-20 min), 100% buffer B (20.01-25 min), 100% buffer A (25.01-30 min). The radioactivity of the eluate was monitored using an in-line NaI radiodetector, digital multichannel analyzer and dedicated software (MetorX B.V.). Elution profiles were analyzed using Empower 3 software (Waters).
Silica gel coated paper
Manufactured by Agilent Technologies
Silica gel-coated paper is a type of laboratory equipment used for chromatographic analysis. It is a thin sheet of paper that has been coated with a layer of silica gel, a porous material commonly used as a stationary phase in various chromatographic techniques. The silica gel coating provides a high surface area and selective adsorption properties, which allow for the separation and analysis of complex mixtures.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using silica gel coated paper
1
Radiolabeled Peptide Characterization
2
Radiolabeling of DTPA-Conjugated Nanobodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
JVZ-007-c-myc-his was incubated with a 5-fold molar excess of p-SCN-Bn-DTPA (Macrocyclics) in 0.1 M sodium carbonate buffer (pH 9.5) for 2.5 h at room temperature. JVZ-007-cys was reduced with 1 mM 2-mercaptoethylamine-HCl in phosphate-buffered saline (PBS), 5 mM ethylenediaminetetraacetic acid for 90 min at 37°C. Reduced JVZ-007-cys was then incubated with 5 mM maleimide-DTPA for 2 h at 37°C. Conjugated Nanobodies were then dialyzed for 3 d in a Slide-A-Lyzer (3.5-kDa cutoff; Life Technologies) against 0.25 M ammonium acetate (NH 4 Ac), pH 5.5.
Nanobody-DTPA conjugates were labeled with 111 InCl 3 (Covidien) in 20 mM sodium acetate, pH 5.0, for 30 min at room temperature. Radioprotectants (3.5 mM ascorbic acid, gentisic acid, and methionine) were used to prevent radiolysis. Labeling efficiency was assessed by instant thin-layer chromatography using silica gel-coated paper (Varian Inc.) and 0.1 M citrate buffer, pH 5.0, as the mobile phase. After incubation, an excess of DTPA (final concentration, 0.15 mM) was added to complex free 111 InCl 3 .
Nanobody-DTPA conjugates were labeled with 111 InCl 3 (Covidien) in 20 mM sodium acetate, pH 5.0, for 30 min at room temperature. Radioprotectants (3.5 mM ascorbic acid, gentisic acid, and methionine) were used to prevent radiolysis. Labeling efficiency was assessed by instant thin-layer chromatography using silica gel-coated paper (Varian Inc.) and 0.1 M citrate buffer, pH 5.0, as the mobile phase. After incubation, an excess of DTPA (final concentration, 0.15 mM) was added to complex free 111 InCl 3 .
3
Efficient Radiolabeling of Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peptides were radiolabeled with 111InCl3 (Curium) in 0.5 M 2-(N-morpholino)ethanesulfonic acid (MES) buffer (twice volume of 111InCl3), pH 5.5, for 10-30 min at 45 °C under metal-free conditions 24 (link). Following incubation, 50 mM ethylenediaminetetraacetic acid (EDTA) was added to a final concentration of 5 mM to chelate unincorporated 111InCl3. Labeling efficiency was determined by instant thin-layer chromatography (ITLC) using silica gel-coated paper (Agilent Technologies) and 0.1 M ammonium acetate containing 0.1 M EDTA, pH 5.5, as the mobile phase. Moreover, radiochemical purity was checked using reverse-phase high performance liquid chromatography (RP-HPLC) on an Agilent 1200 system (Agilent Technologies) with an in-line radiodetector. A C18 column (5 µm, 4.6 × 250 mm; HiChrom) was used at a flow rate of 1 ml/min with the following buffer system: buffer A, triethylammonium acetate (10 mM, pH 7); buffer B, 100% methanol; and a gradient of 97% to 0% buffer A (35 min). Peptides were purified by a Sep-Pak C18 light cartridge (Waters) and eluted from the cartridge with 50% ethanol in water.
4
Radiolabeling Peptides with Indium-111
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peptides were labeled under metal-free conditions with 111InCl3 (Curium) in 0.5 M 2-(N-morpholino)ethanesulfonic acid (MES) buffer (pH 5.5, twice volume of 111InCl3) or sodium acetate buffer (NaOAc in 0.04 M acetic acid solution, pH 4.5). Labeling was performed at 45 °C for 10 min [26 (link)]. To chelate unincorporated 111InCl3, ethylenediamine-tetraacetic acid (EDTA, 50 mM) was added to a final concentration of 5 mM after the incubation.
Radiochemical yield (RCY) was determined by instant thin-layer chromatography (ITLC) using silica gel-coated paper (Agilent Technologies) and 0.1 M ammonium acetate containing 0.1 M EDTA, pH 5.5, as the mobile phase. In addition, RCY was determined using RP-HPLC on an Agilent 1200 system (Agilent Technologies) with an in-line radiodetector (Elysisa-Raytest). A reversed-phase C18 column (5 µm, 4.6 × 250 mm; HiChrom) was used at a flow rate of 1 ml/min. We used the following buffer system: buffer A, triethylammonium acetate (TEAA, 10 mM, pH 7); buffer B, 100% methanol; and a gradient of 97 to 0% buffer A (35 min). Peptides were purified by a Sep-Pak C18 light cartridge (Waters) and eluted from the cartridge with 50% ethanol in water.
Radiochemical yield (RCY) was determined by instant thin-layer chromatography (ITLC) using silica gel-coated paper (Agilent Technologies) and 0.1 M ammonium acetate containing 0.1 M EDTA, pH 5.5, as the mobile phase. In addition, RCY was determined using RP-HPLC on an Agilent 1200 system (Agilent Technologies) with an in-line radiodetector (Elysisa-Raytest). A reversed-phase C18 column (5 µm, 4.6 × 250 mm; HiChrom) was used at a flow rate of 1 ml/min. We used the following buffer system: buffer A, triethylammonium acetate (TEAA, 10 mM, pH 7); buffer B, 100% methanol; and a gradient of 97 to 0% buffer A (35 min). Peptides were purified by a Sep-Pak C18 light cartridge (Waters) and eluted from the cartridge with 50% ethanol in water.
5
Radiolabeling of PSMA-Targeted Tracer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PSMA I&T (Fig. 1 ), a DOTAGA-chelated urea-based PSMA inhibitor, was synthetized as described previously 25 (link) and kindly provided by the Technical University Munich. PSMA I&T was radiolabeled with 111InCl3 (Mallinckrodt BV, Petten, The Netherlands) in 0.5 M 2-(N-morpholino)ethanesulfonic acid (MES) buffer (twice volume of 111InCl3), pH 5.5, for 10 min at 95 °C under metal-free conditions 29 (link). After incubation, 4 mM diethylenetriaminepentaacetic acid (DTPA) was added to a final concentration of 0.2 mM to complex free 111InCl3. Labeling efficiency was determined by instant thin-layer chromatography (ITLC) using silica gel coated paper (Agilent Technologies, Palo Alto, CA) and 0.1 M ammonium acetate containing 0.1 M ethylenediaminetetraacetic acid (EDTA), pH 5.5, as the mobile phase. Labeling efficiency exceeded 95% in all experiments.
6
Radiolabeling of PSMA Ligands with 111In
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PSMA ligands (1 µg/labeling; specific activity 5 MBq/µg) were radiolabeled with 5 MBq 111InCl3 (curium) in 0.5 M 2-(N-morpholino)ethanesulfonic acid buffer (twice volume of 111InCl3), pH 5.5, at 45 °C for 30 min under metal-free conditions [20 (link)]. After incubation, 50 mM ethylenediaminetetraacetic acid (EDTA) was added to a final concentration of 5 mM to chelate unincorporated 111InCl3. Labeling efficiency was determined by instant thin-layer chromatography (ITLC) using silica gel–coated paper (Agilent Technologies) and 0.1 M ammonium acetate containing 0.1 M EDTA, pH 5.5, as the mobile phase. To determine the effects of 111In labeling on tPDT efficacy, an in vitro tPDT assay was performed with and without radiolabeling of PSMA-N064 (3 and 30 nM, 5 MBq 111In).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!