All solvents and chemicals were of reagent grade quality, purchased from Sigma-Aldrich (St. Louis, MO), and used without further purification unless otherwise noted. G5 PAMAM dendrimer was purchased from Dendritech (Midland, MI). Monomethyl-PEG11-NHS (PEG-NHS) ester was purchased from ChemPep Inc. 3-(4-(2-azidoethoxy) phenyl) propanoic acid was used from previous studies. 2,2′,2′′-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetic acid (DOTA-NHS) was purchased from Macrocyclics (Dallas, TX). AF647-azide was purchased from Life Technologies of Thermo Fisher Scientific (Grand Island, NY). Trastuzumab was obtained from Genetech (San Francisco, CA).
Dota nhs
Manufactured by Macrocyclics
Sourced in United States
DOTA-NHS is a bifunctional chelator that can be used for the conjugation of molecules to proteins. It contains a DOTA chelator for the complexation of metal ions and an N-hydroxysuccinimide (NHS) ester for coupling to primary amines on biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Maleimide dota, by Macrocyclics (2 mentions)
Alexa fluor 647 nhs ester, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 647 c2 maleimide, by Thermo Fisher Scientific (2 mentions)
Trastuzumab, by Gene Tech (1 mentions)
Fetal bovine albumin, by Thermo Fisher Scientific (1 mentions)
Pd 10 desalting column, by Cytiva (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Milli q plus 185 water purification system, by Merck Group (1 mentions)
Herceptin, by Gene Tech (1 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)
Amicon ultra 4 centrifugal filter unit, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Xtt assay, by Roche (1 mentions)
Hepes buffer, by Thermo Fisher Scientific (1 mentions)
35 mm glass bottom culture dishes, by MatTek (1 mentions)
A549 cells ccl 185, by American Type Culture Collection (1 mentions)
Rucaparib, by Selleck Chemicals (1 mentions)
Gemcitabine, by Pfizer (1 mentions)
22 protocols using dota nhs
1
Synthesis of Dendrimer-PEG Conjugate
2
In-111 Radiolabeling and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
111In was produced in Radiation Application Research School, Karaj, Iran, by 112Cd (p, 2n) 111In reaction. DOTA-NHS was purchased from Macrocyclics (NJ, USA). Fetal Bovine Albumin, RPMI-1640 medium, and L-Glutamine were bought from Gibco Co. (Dublin, Ireland). PD10 De-salting column was inquired from Amersham Pharmacia Biotech; additional chemicals were purchased from Sigma Chemical Co. (MO, USA). Sprague-Dawley rats were obtained from Pasteur Institute (Tehran, Iran). A Bioscan AR-2000 radio thin-layer chromatography (TLC) scanner instrument (Bioscan, Paris, France) was used for Radio-chromatography purposes. A p-type coaxial high-purity germanium (HPGe) detector (model: EGPC 80–200R) coupled with a multichannel analyzer card system and a dose calibrator ISOMED 1010 (Dresden, Germany) were utilized for the measurement of the activity. Calculations were carried out based on the 245 keV peak for 111In. The United Kingdom Biological Council's Guidelines on the Use of the Living Animals in Scientific Investigations, 2nd edition was used to determine the framework of animal experiments. Achieved results are displayed as mean ± standard deviation (mean ± standard deviation), and Student's t-test was used to compare the data based on statistical significance defined as P < 0.05.
3
Chemoselective DOTA-NHS Peptide Conjugation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 17
DOTA-NHS (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid N-hydroxy-succinimide ester) was purchased from Macrocyclics, Inc. Dallas, USA. Conjugation of DOTA-NHS to TCO-peptide 9 employed a two-step procedure as illustrated in Scheme 14.
In the first step, TCO-peptide 9 was dissolved in the conjugation buffer (phosphate buffered saline, PBS, with 5 mM EDTA pH 7.0) at 1 mM. The reaction mixtures were incubated for overnight at room temperature. In the second step, the DOTA-NHS ester was added to the incubated solution at 100 mM final concentration (1:100 molar ratio or 1:20 equivalent ratio). Since the DOTA-NHS ester was acidic because of containing TFA, the pH of the solution was adjusted to 8.0 in order to activate the NHS ester-NH2 coupling reaction. The reaction mixtures were incubated overnight at room temperature.
According to the data in
4
Dendrimer-based Targeted Cancer Therapy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All solvents and chemicals were of reagent grade quality, purchased from Sigma-Aldrich (St. Louis, MO), and used without further purification unless otherwise noted. G5 PAMAM dendrimer was purchased from Dendritech (Midland, MI). Monomethyl-PEG11-NHS (PEG-NHS) ester was purchased from ChemPep Inc. 3-(4-(2-azidoethoxy)phenyl) propanoic acid was used from previous studies [37 (link)]. 2,2′,2′′-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane- 1,4,7-triyl) triacetic acid (DOTA-NHS) was purchased from Macrocyclics (Dallas, TX). AF647-azide was procured through Life Technologies of Thermo Fisher Scientific (Grand Island, NY). Herceptin was obtained from Genetech (San Francisco, CA) Water used in all experiments was purified using a Milli-Q Plus 185 water purification system (Millipore, Bedford, MA) with resistivity higher than 18 MΩ cm; PBS and Amicon® Ultra-4 centrifugal filter units (MWCO = 10,000 or 100,000) were purchased from Millipore. 35 mm glass bottom culture dishes were purchased from MatTek Corporation, the ProLong Gold Antifade Reagent with DAPI from Life Technologies, and the XTT assay was bought from Roche. DMEM, HEPES buffer and Trypsin-EDTA, all were purchased from Gibco. SKBR-3 cells (HTB-30) and A549 cells (CCL-185) were purchased from American Type Culture Collection.
5
Combination Chemo and RIT in Tumor-Bearing Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumour-bearing mice were treated intravenously with 50 mg/kg gemcitabine (Hospira, Melbourne, VIC, Australia) on days 1 and 2 and 2.5 mg/kg cisplatin (Hospira) on day 1. DAB4 and Sal5 were conjugated to the bi-functional chelator DOTA-NHS (Macrocyclics, Dallas, TX, USA) as previously described
[11 (link)] and radiolabeled with 177Lu (PerkinElmer, Waltham, MA, USA). Radioimmunoconjugates were administered intravenously on day 3. The specific activity of radioimmunoconjugates ranged from 95 to 130 MBq/mg with >97% incorporation of 177Lu as determined by instant thin layer chromatography. The PARPi inhibitor Rucaparib (AG-014699; Selleck Chemicals, Houston, TX, USA) was diluted in 5% D-glucose in PBS for intraperitoneal injection at 1 or 2 mg/kg and administered daily on days 1 to 5, 30 min before chemotherapy or RIT. For in vivo antibody binding analysis, DAB4 was biotinylated with EZ-Link NHS-Biotin (Thermo Fisher) following manufacturer’s instructions. One hundred micrograms of biotin-DAB4 was administered 24 h after chemotherapy.
[11 (link)] and radiolabeled with 177Lu (PerkinElmer, Waltham, MA, USA). Radioimmunoconjugates were administered intravenously on day 3. The specific activity of radioimmunoconjugates ranged from 95 to 130 MBq/mg with >97% incorporation of 177Lu as determined by instant thin layer chromatography. The PARPi inhibitor Rucaparib (AG-014699; Selleck Chemicals, Houston, TX, USA) was diluted in 5% D-glucose in PBS for intraperitoneal injection at 1 or 2 mg/kg and administered daily on days 1 to 5, 30 min before chemotherapy or RIT. For in vivo antibody binding analysis, DAB4 was biotinylated with EZ-Link NHS-Biotin (Thermo Fisher) following manufacturer’s instructions. One hundred micrograms of biotin-DAB4 was administered 24 h after chemotherapy.
6
Synthesis and Purification of DOTA-GSAO
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Diethylene triamine pentaacetic acid GSAO (DTPA-GSAO) was synthesized as previously described [17 (link)]. Stocks of this were diluted to a concentration of 1 mg/mL and 46 μg aliquots stored at − 20 °C in nitrogen filled glass vials.
Preparation of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid GSAO (DOTA-GSAO) was undertaken by dissolving GSAO (155 mg, 0.28 mmol) in 20 mL of argon purged water. While stirring under argon, the pH of the solution was adjusted to 7.2 by slow addition of 0.1 M NaHCO3. DOTA-NHS (258 mg, 0.34 mmol) (Macrocyclics) was added in two equal portions with a pH adjustment to 7.0 after each addition using 0.1 M NaHCO3. The reaction mixture was stirred for 30 min at room temperature following the final addition. The crude product solution was purified on a Biotage RP18 Cartridge (YMC-GEL, ODS-AQ, 120-S50) with water-acetonitrile gradient as eluant to yield 99% pure product (105 mg, 40% yield) upon lyophilization.
Preparation of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid GSAO (DOTA-GSAO) was undertaken by dissolving GSAO (155 mg, 0.28 mmol) in 20 mL of argon purged water. While stirring under argon, the pH of the solution was adjusted to 7.2 by slow addition of 0.1 M NaHCO3. DOTA-NHS (258 mg, 0.34 mmol) (Macrocyclics) was added in two equal portions with a pH adjustment to 7.0 after each addition using 0.1 M NaHCO3. The reaction mixture was stirred for 30 min at room temperature following the final addition. The crude product solution was purified on a Biotage RP18 Cartridge (YMC-GEL, ODS-AQ, 120-S50) with water-acetonitrile gradient as eluant to yield 99% pure product (105 mg, 40% yield) upon lyophilization.
7
Radiolabeling of DOTA-conjugated Dia12.3 Antibody
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A 20-fold excess of DOTA-NHS (220 nmol, Macrocyclics, TX) in 4.5μl of H2O was added to 1.65 mg (11 nmol) of Dia12.3 antibody in 2.5 ml of PBS1X, and the pH was adjusted to 7.36 with 0.1N NaOH. The reaction solution was rotated at the RT under Argon overnight. Then the reaction mixture was dialyzed against the PBS 1X buffer with five times and sterile filtered. Dota conjugation was confirmed by Instant thin layer chromatogram (ITLC) performed by the Radiopharmacy Department at City of Hope. One hundred μg of DOTA-DIA 12.3 was radiolabeled with 10 mCi of 64CuCl2 in 0.1 M HCl at pH 4.5 by adjusting the pH with 0.2M ammonium acetate. The final product purified by Size-Exclusion Chromatography by the Radiopharmacy Department of City of Hope.
8
DOTA-NHS Peptide Conjugation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 18
DOTA-NHS (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid N-hydroxy-succinimide ester) was purchased from Macrocyclics, Inc. Dallas, USA. Conjugation of DOTA-NHS to TCO-peptide 9 employed a two-step procedure as illustrated in Scheme 9. In the first step, TCO-peptide 9 was dissolved in the conjugation buffer (phosphate buffered saline, PBS, with 5 mM EDTA pH 7.0) at 1 mM. The reaction mixtures were incubated for overnight at room temperature. In the second step, the DOTA-NHS ester was added to the incubated solution at 100 mM final concentration (1:100 molar ratio or 1:20 equivalent ratio). Since the DOTA-NHS ester was acidic because of containing TFA, the pH of the solution was adjusted to 8.0 in order to activate the NHS ester-NH2 coupling reaction. The reaction mixtures were incubated overnight at room temperature.
According to the data in
9
Radiolabeling and Imaging of Monoclonal Antibody
10
DOTA-NHS Conjugation to TCO-Peptide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 10
DOTA-NHS (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid N-hydroxy-succinimide ester) was purchased from Macrocyclics, Inc. Dallas, USA. Conjugation of DOTA-NHS to TCO-peptide 9 employed a two-step procedure as illustrated in Scheme 4. In the first step, TCO-peptide 9 was dissolved in the conjugation buffer (phosphate buffered saline, PBS, with 5 mM EDTA pH 7.0) at 1 mM. The reaction mixtures were incubated for overnight at room temperature. In the second step, the DOTA-NHS ester was added to the incubated solution at 100 mM final concentration (1:100 molar ratio or 1:20 equivalent ratio). Since the DOTA-NHS ester was acidic because of containing TFA, the pH of the solution was adjusted to 8.0 in order to activate the NHS ester-NH2 coupling reaction. The reaction mixtures were incubated overnight at room temperature.
According to the data in
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!