1×106 LS174T cells expressing firefly luciferase were delivered intraperitoneally (i.p.). 1×106 NK-92MI cells were labeled using Cy5.5 NHS ester (Lumiprobe Corporation, Hannover, Germany) for 30 min before injection, according to the manufacturer’s instructions, and delivered either by intravenous (i.v.) or i.p. injections seven days post-tumor cell injection. Bioluminescence and fluorescence were determined using an in vivo imaging system (IVIS200 system, Perkin Elmer, Waltham, MA, USA). At study termination, mice were sacrificed and samples of tumors, serums, peritoneal washes, spleens, and livers were collected for further analysis.
Cy5.5 nhs ester
Manufactured by Lumiprobe
Sourced in United States
Cy5.5 NHS ester is a fluorescent labeling reagent used in biomedical research. It is a cyanine dye that can be used to label biomolecules such as proteins, peptides, and nucleic acids. The NHS (N-hydroxysuccinimide) ester group allows the dye to react with primary amines, forming a stable amide bond.
Automatically generated - may contain errors
Lab products found in correlation
Irdye800cw maleimide, by LI COR (2 mentions)
Origin 6, by OriginLab (2 mentions)
Pd 10 desalting column, by GE Healthcare (2 mentions)
Cy5 nhs ester, by Lumiprobe (2 mentions)
Ivis 200 system, by PerkinElmer (1 mentions)
Pooled human saliva, by Lee Biosolutions (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Ammonium bicarbonate, by Merck Group (1 mentions)
Leupeptin, by Merck Group (1 mentions)
Actin tracker green 488, by Beyotime (1 mentions)
Saporin, by Merck Group (1 mentions)
Bca assay kit, by Beyotime (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Sangon (1 mentions)
2 n morpholino ethanesulfonic acid mes, by Merck Group (1 mentions)
Goat anti mouse igg h l, by Abcam (1 mentions)
Milli q system, by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
24 protocols using cy5.5 nhs ester
1
Tumor Targeting with NK Cells
2
Peptide-Based Fluorescent Salivary Assay
3
Chondrocyte Imaging with Copper-based Dyes
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Chondrocytes were seeded in confocal dishes (1 × 105 cells), and then incubated with Cy5.5-Cu1.5MH or Cy5.5-Cu6MH (20 μg/ml) in the dark for 24 h. After incubation, chondrocytes were washed with PBS 3 times and then fixed with paraformaldehyde (4% PFA, Biosharp, China) for 10 min. Later, the cytoskeleton was stained with the actin-tracker green-488 (Beyotime Biotechnology, China) following the protocols, and the nuclei was stained with 4, 6-diamidino-2-phenyindole dilactate (DAPI, Beyotime Biotechnology, China) for 10 min before PBS washing 3 times. Finally, the images were captured with a confocal scanning microscope (Leica, Germany), and the corresponding fluorescence intensity was quantified by ImageJ. Specifically, Cy5.5-Cu1.5MH and Cy5.5-Cu6MH were prepared by the following procedures. In detail, 50 mg of Cu1.5MH or Cu6MH was dispersed in DMSO followed by the addition of 50 mg/ml Cy5.5 N-hydroxysuccinimide ester (Cy5.5 NHS ester, Lumiprobe, USA) overnight. The final product was collected by vacuum drying after methanol washing. Besides, chondrocytes were incubated with Cu6MH (20 μg/ml) for 12 h, and chondrocytes were also embedded in paraffin and cut into slice for observation by TEM.
4
Fluorescent Peptide Synthesis and Characterization
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The fluorescently-labeled peptides were synthesized using standard solid phase Fmoc chemistry, as described previously [25 (link)]. 5-FITC was obtained from Sigma Aldrich. Cy5 and Cy5.5 NHS ester were obtained from Lumiprobe. IRDye800CW maleimide was obtained from LI-COR Biosciences. Fluorescence emission was collected with a fiber coupled spectrophotometer (Ocean Optics) using excitation at λex =488, 660, and 785 nm. The spectra were plotted using Origin 6.1 software (Origin Lab Corp).
5
Fluorescent Peptide Synthesis and Characterization
6
Dendrimer-Based Targeted Drug Delivery
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Generation 5 amine-terminated polyamidoamine dendrimer (G5-NH2) with an ethylenediamine core was obtained from Dendritech Inc. (Midland, MI, USA). 4-(Bromomethyl)phenylboronic acid was purchased from Meryer Chemical Technology Co., Ltd. (Shanghai, China). Saporin and PASP sodium (2000–11000 Da) were purchased from Sigma-Aldrich (St Louis, MO). Cy5.5 NHS ester was purchased from Lumiprobe (USA). Fluorescein isothiocyanate (FITC) was bought from Macklin Biochemical Co., Ltd. (Shanghai, China). Methanol and dimethyl sulfoxide (DMSO) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Sangon Biotech. (Shanghai, China). BCA assay kit were purchased from Beyotime (Jiangsu, China).
7
Synthesis and Characterization of Targeted PSMA Nanoparticles
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Triton X-100 (TX-100), cyclohexane and n-hexyl alcohol were obtained from Alfa Aesar (Ward Hill, MA, USA). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS), tetraethyl orthosilicate (TEOS), 3-aminopropyltrimethoxysilane (APTES) and 2-N-morpholino-ethanesulfonic acid (MES) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cy5.5-NHS ester was purchased from Lumiprobe Corporation (Maryland, USA). Saline-PEG-COOH was purchased from Ponsurebio Co., Ltd. (Shanghai, China). Ammonia solution (25–28%), acetone and anhydrous ethanol (99.5 %) were purchased from Chuandong Chemical Co., Ltd. (Chongqing, China). Anti-PSMA antibody (YPSMA-1), goat anti-mouse IgG and goat anti-mouse IgG H&L (FITC) were purchased by Abcam (Cambridge, UK). Gadodiamide (OmniscanTM) was obtained from GE Healthcare (Cork, Ireland). Deionized water was provided by a Milli-Q system from Millipore (Millipore, USA). All chemicals were reagent grade.
8
Synthesis and Characterization of PF Formulation
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PF (purity >98%) was purchased from Chengdu Biopurify Phytochemicals Ltd. (Chengdu, People’s Republic of China). Lipoid S 80 (phosphatidylcholine containing 76% of soybean lecithin) was obtained from Lipoid GmbH (Ludwigshafen, Germany). Cy5.5-NHS ester was purchased from Lumiprobe Corporation (Hallandale Beach, FL, USA). S. tuberculata was obtained from Bozhou Changfu Pharmaceutical Ltd. (Anhui, People’s Republic of China). PF and salidroside standards (purity >99.8%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, People’s Republic of China). Liquid chromatography-grade methanol and acetonitrile were purchased from Merck (Darmstadt, Germany). Liquid chromatography-grade ammonium acetate was purchased from Sigma-Aldrich (St Louis, MO, USA). Other chemicals were obtained from Sinopharm Chemical Reagent (Shanghai, People’s Republic of China) and were of analytical grade.
9
Fluorescent Lipid Conjugate Synthesis
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Cy5.5 NHS ester was acquired from Lumiprobe (Hunt Valley, MD, USA). Bovine serum albumin (BSA) was procured from Amresco (Solon, OH, USA). Cholesterol, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000) were purchased from Avanti Polar Lipids (Alabaster, AL, USA).
10
Synthesis and Characterization of Cy5.5-Labeled ZD2 Peptide
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ZD2 peptide (sequence: Thr-Val-Arg-Thr-Ser-Ala-Asp) was synthesised in solid phase using standard Fmoc-chemistry. After Fmoc removal with 10% piperidine, Fmoc-9-amino-4,7-dioxanonanoic acid (ChemPep, Inc., Wellington, FL, USA) was added to the peptide sequence. After Fmoc removal with 10% piperidine, the resin was washed with DMF/DCM and air-dried, and 10 mg of the dried resin was swelled in DCM for 1 h, followed by reaction with 3 mg Cy5.5-NHS ester (Lumiprobe Corporation, Hallandale Beach, FL, USA) in presence of 5 μL DIPEA. Reaction was stirred overnight at room temperature. Excess Cy5.5-NHS ester was removed by filtration and washing with DMF/DCM 10 mL three times. Peptides were cleaved off resin using TIPS, and precipitated in cold ether. The products were separated from ether by centrifugation at 4000×g. The final product was characterised by MALDI-TOF mass spectrometry ([M+1]+: m/z = 1473.76 (obsd); 1472.78 (calc.)). The product was lyophilised and reconstituted in 500 μL PBS. The concentration of the solution was characterised by measuring absorbance at 650 nm.
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