Endotoxin-free F-TRAP was subsequently diluted in endotoxin-free 50 mM carbonate/bicarbonate buffer, pH 8.5, to 1 mg/mL and conjugated to the NIR fluorochrome VivoTag-S 750 (PerkinElmer) at room temperature for 1 h using end-over-end mixing (Labquake Rotisserie, Barnstead Thermolyne). Nonconjugated fluorochrome was removed using an Amicon Ultra-15 centrifugal filter 3 kDa MWCO, while buffer exchanging fluorochrome-conjugated F-TRAP (NIR-F-TRAP) into endotoxin-free 1× PBS, pH 7.4, and concentrating to 13.40 mg/mL.
Vivotag s 750
Manufactured by PerkinElmer
Sourced in United States
VivoTag-S 750 is a fluorescent dye that can be used for in vivo imaging. It exhibits high quantum yield and stability, making it suitable for long-term imaging studies. The dye has an absorption maximum at 750 nm and an emission maximum at 780 nm, allowing for deep tissue imaging with minimal autofluorescence interference.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey clx imaging system, by LI COR (3 mentions)
Lysing matrix a tube, by MP Biomedicals (2 mentions)
Fastprep 24 homogenizer, by MP Biomedicals (2 mentions)
Ivis spectrum ct imaging system, by PerkinElmer (2 mentions)
Vivotag s 680, by PerkinElmer (1 mentions)
Capillary tubes, by Avantor (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Mmpsense 645 fast, by PerkinElmer (1 mentions)
Angiosense 680, by PerkinElmer (1 mentions)
Fmt 4000 system, by PerkinElmer (1 mentions)
Inveon pet ct, by Siemens (1 mentions)
Cut off centrifugal filter, by Merck Group (1 mentions)
Vivotag 750, by PerkinElmer (1 mentions)
Vivotag s 750 mal, by PerkinElmer (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Spark multimode microplate reader, by Tecan (1 mentions)
Zetasizer nano, by Malvern Panalytical (1 mentions)
8 arm peg amine, by JenKem Technology (1 mentions)
Rmil 2, by R&D Systems (1 mentions)
Rmil 15, by R&D Systems (1 mentions)
11 protocols using vivotag s 750
1
Fluorescent TRAP Conjugation Protocol
2
Dual-Targeted Liposomal Nanoparticle Synthesis
3
Characterizing Antibody-GS Conjugate Pharmacokinetics
4
Multimodal Imaging of Vascular Targeting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood clearance, organ distribution and preliminary imaging studies used IVIS Spectrum (Caliper LifeSciences) together with micro-CT (Inveon PET-CT, Siemens). Subsequently we used an FMT 4000 system (PerkinElmer, Massachusetts USA) equipped with four lasers. Animals were injected with a VivoTag-S 750-MAL-conjugated antibody, as well as in selected experiments with MMPSense-645-FAST (PerkinElmer, MA) to evaluate MMP activity or AngioSense 680 (PerkinElmer, MA) to identify intravascular space. Contrast-enhanced high resolution CT localized the aortic root and major arteries in the neck and thorax to guide the placement of the region of interest (ROI) for the fluorescence activity map concomitantly obtained by FMT. Fluorescence and CT image fusion relied on fiducial markers present on the imaging cartridge, and used Amide 1.0.4 (Sourceforge.net).
5
PVX Modification with Fluorescent Dyes and PEG
6
In Vivo Nanosensor Biodistribution Monitoring
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice bearing skin grafts were administered with nanosensors (20 μg) or peptides (10 nmol) labelled with VivoTag S-750 (VT750; PerkinElmer). Nanosensors and peptides were labeled with VT750 (3:1 molar ratio) using NHS chemistry per manufacturer’s protocol. For organ biodistribution, whole mice were imaged with IVIS Spectrum CT Imaging System (PerkinElmer) while excised organs were imaged with Odyssey CLx Imaging System (LI-COR) 24 hours after administration. For urine pharmacokinetics, whole mice were imaged with IVIS Spectrum CT Imaging System (PerkinElmer) at 90 minutes post administration. To track cleaved fragments after in vivo GzmB cleavage, on day 7 post-transplant, VT750-labelled activity nanosensors (10 nmol by peptides) were administered to skin graft mice. At 90-minute post administration, whole mouse was imaged with IVIS Spectrum CT Imaging System (PerkinElmer) to analyze the extend of fluorescence from the bladder. Major organs were then excised and homogenized using Lysing Matrix A Tubes and FastPrep24 Homogenizer (MP Biomedicals). Tissue homogenates were transferred to a 96-well plate and then imaged with Odyssey CLx Imaging System (LI-COR).
7
Peptide-Targeted Fluorescent Probes for TBI
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following targeting peptides were synthesized
by Lifetein (Somerset, NJ): PGpep (X(FAM)-CAQK), CIVpep (KLWVLPKGG-K(FAM)-GGC),
FIBpep (X(FAM)-CREKA), HApep (X(FAM)-CSTMMSRSHKTRSHHV), and
RGD (X(FAM)-CRGD). Calpain substrate FRET peptide (QSY21-QEVYGAMP-K(Cy5)-PEG2-GC-NH2)
was synthesized by CPC Scientific, Inc. (Sunnyvale, CA). X stands
for 6-aminocaproic acid, and PEG2 stands for poly(ethylene glycol).
The 40 kDa 8-arm PEG maleimide (tripentaerythritol) was purchased
from Jenkem Technology (Beijing, China). For ECM-targeting peptide
screening, PEG maleimide was batch-reacted with 1 mol ofl -cysteine-functionalized VivoTag S-750 (PerkinElmer, Boston, MA).
The reaction was then split to ensure matched VivoTag 750 modification
for each conjugate and reacted with 8.4 mol equiv of each ECM-targeting
peptide followed by quenching with an excess ofl -cysteine.
For HA-targeting TBI-ABNs: The 8-arm PEG maleimide was batch reacted
with 1 mol equiv of calpain substrate FRET peptide, split, and reacted
with 0 mol equiv (nontargeting), 4 mol equiv (moderate-targeting),
or 8.4 mol equiv (high-targeting) of HApep in the presence of 50 mM
triethylamine (TEA), then quenched with an excess ofl -cysteine.
All conjugates were dialyzed with water, and final concentrations
were determined by absorbance of VivoTag 750, FAM, or Cy5 using a
Spark multimode microplate reader (Tecan Trading AG, Switzerland).
by Lifetein (Somerset, NJ): PGpep (X(FAM)-CAQK), CIVpep (KLWVLPKGG-K(FAM)-GGC),
FIBpep (X(FAM)-CREKA), HApep (X(FAM)-CSTMMSRSHKTRSHHV), and
RGD (X(FAM)-CRGD). Calpain substrate FRET peptide (QSY21-QEVYGAMP-K(Cy5)-PEG2-GC-NH2)
was synthesized by CPC Scientific, Inc. (Sunnyvale, CA). X stands
for 6-aminocaproic acid, and PEG2 stands for poly(ethylene glycol).
The 40 kDa 8-arm PEG maleimide (tripentaerythritol) was purchased
from Jenkem Technology (Beijing, China). For ECM-targeting peptide
screening, PEG maleimide was batch-reacted with 1 mol of
The reaction was then split to ensure matched VivoTag 750 modification
for each conjugate and reacted with 8.4 mol equiv of each ECM-targeting
peptide followed by quenching with an excess of
For HA-targeting TBI-ABNs: The 8-arm PEG maleimide was batch reacted
with 1 mol equiv of calpain substrate FRET peptide, split, and reacted
with 0 mol equiv (nontargeting), 4 mol equiv (moderate-targeting),
or 8.4 mol equiv (high-targeting) of HApep in the presence of 50 mM
triethylamine (TEA), then quenched with an excess of
All conjugates were dialyzed with water, and final concentrations
were determined by absorbance of VivoTag 750, FAM, or Cy5 using a
Spark multimode microplate reader (Tecan Trading AG, Switzerland).
8
In Vivo Nanosensor Biodistribution Monitoring
9
Fluorescent Antibody Conjugation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Conjugation of 78Fc or control human IgG (Genscript) with VivoTag-S 750 and VivoTag-S 750MAL (Perkin Elmer) was performed according to the manufacturer's instructions. Briefly, 1 mL of a 1 mg/mL antibody solution in conjugation buffer (50mM carbonate/bicarbonate buffer, pH8.5) was mixed with 30μL of VivoTag-S 750 at a concentration of 10mg/mL in Dimethylsulfoxide (Sigma) and then incubated at room temperature for 1 h. After dialysis in 1×PBS, the reaction mixture was filtered through a 0.2 μm syringe filter. Aliquots of labeled protein were stored at 4 °C in the dark until ready to analyze.
10
Calpain Substrate Peptide Conjugation
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!