Cy5.5 nhs

Manufactured by Lumiprobe

Sourced in United States

Cy5.5-NHS is a fluorescent dye used in various biological and biochemical applications. It is a water-soluble, succinimidyl ester derivative of the Cy5.5 fluorophore, which is designed for covalent labeling of proteins and other biomolecules containing primary amine groups. Cy5.5-NHS exhibits excitation and emission wavelengths suitable for detection in the far-red region of the visible spectrum.

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Lab products found in correlation

Human serum albumin (hsa), by Solarbio (1 mentions) D luciferin potassium salt, by Merck Group (1 mentions) Balb c nude mice, by Charles River Laboratories (1 mentions) Antimycin a, by Merck Group (1 mentions) Carbonyl cyanide p, by Merck Group (1 mentions) Chloroquine diphosphate cq, by Merck Group (1 mentions) Low melting point agarose, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions) Hydrazine hydrate, by J&K Scientific (1 mentions) α sma, by Abways (1 mentions) Gemcitabine hydrochloride, by Meilun (1 mentions) Delta prep 4000, by Waters Corporation (1 mentions) Voyager workstation, by Thermo Fisher Scientific (1 mentions) Tcs sp8, by Leica (1 mentions) Airfuge air driven ultracentrifuge, by Beckman Coulter (1 mentions) Amicon ultra, by Merck Group (1 mentions) Cy3 nhs, by Lumiprobe (1 mentions) Thiazolyl blue tetrazolium bromide mtt, by Thermo Fisher Scientific (1 mentions) 1 3 dimethylaminopropyl 3 ethylcarbodiimide hydrochloride edc, by Thermo Fisher Scientific (1 mentions) Phosphotungstic acid pta, by Electron Microscopy Sciences (1 mentions)

Spelling variants of this product

Cy5.5 NHS ester (24 citations) Cy5-NHS (21 citations) Sulfo-Cy5.5-NHS ester (4 citations) Cyanine5.5 N-hydroxysuccinimide ester (Cy5.5-NHS) (3 citations) Sulfo-Cyanine 5 (Cy5) NHS ester (2 citations) Cy5.5 N-hydroxysuccinimide (Cy5.5-NHS) ester (2 citations) Cy5.5 NHS dye (2 citations) Cyanine5.5 NHS ester (Cy5.5) (2 citations)

9 protocols using cy5.5 nhs

Recombinant Ferritin Purification and Labeling

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Ferritin
was expressed in Escherichia coli (E. coli) cells and purified by gel filtration. Human
serum albumin was purchased
from Solarbio. NHS-PEG-MAL, disulfiram, CuCl₂, Na₂CO₃, and all buffers were purchased from Aladdin. DMSO
was purchased from Sigma-Aldrich. Cy5.5-NHS was purchased from Lumiprobe. d-Luciferin potassium salt was purchased from Sigma-Aldrich.
All Balb/c nude mice were purchased from Charles River. Animal experiments
were performed in accordance with the guidelines from the IACUC of
Peking University Health Science Center, Beijing, China.

Xu N., Yang Y.F., Chen L, & Lin J. (2020). A Ferritin–Albumin–Cu Nanoparticle that Efficaciously Delivers Copper(II) Ions to a Tumor and Improves the Therapeutic Efficacy of Disulfiram. ACS Omega, 5(18), 10415-10422.

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Adipogenesis Assay with Adipogenic Modulators

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All chemicals, including sodium
SC (S9637), oligomycin, carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone
(FCCP), rotenone, antimycin-A, triiodo-l-thyronine (T3),
indomethacin, 3-isobutyl-1-methylxanthine (IBMX), dexamethasone, and
insulin, were obtained from Sigma-Aldrich (Saint Louis, MO, USA).
HA (MW 8000–15000) was purchased from CARBOSYNTH (Berkshire,
UK). Fluorescent dye Cy5.5-NHS was purchased from the Lumiprobe Corporation
(Hunt Valley, Maryland, USA). Bone morphogenetic protein 7 (BMP7)
was purchased from R&D Systems (Minneapolis, MN, USA). Primary
antimouse UCP1 (#GTX112784) and F4/80 (#ab6640) antibodies were purchased
from GeneTex (Irvine, CA, USA) and Abcam (Cambridge, UK), respectively.

Liao F.H., Yao C.N., Chen S.P., Wu T.H, & Lin S.Y. (2022). Transdermal Delivery of Succinate Accelerates Energy Dissipation of Brown Adipocytes to Reduce Remote Fat Accumulation. Molecular Pharmaceutics, 19(11), 4299-4310.

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Gemcitabine and Chloroquine Combination Therapy

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Gemcitabine hydrochloride was purchased from Meilun Biological Technology (Dalian, China). Chloroquine diphosphate (CQ) and phosphate-PEG₅₀₀₀-NH₂ (PEG_5K) were purchased from Sigma-Aldrich (USA). The third-generation dendrimer poly-lysine (DGL-G3) was purchased from Colcom (France). Succinic anhydride (SA), 6-phosphonohexanoic acid (6PA), 2-propyl-3-methyl maleic anhydride (CDM) and 4-dimethylaminopyridine (DMAP) were purchased from TCI Chemicals (Shanghai, China). N-hydroxy-succinimide (NHS), 1-[3-(dimethylamino) propyl]-3-ethylcarbodiimide hydrochloride (EDC), ultra-dry N-N-dimethylformamide (DMF), ultra-dry dimethyl sulfoxide (DMSO) and hydrazine hydrate were obtained from J&K Scientific (Beijing, China). Cy5.5-NHS was purchased from Lumiprobe (USA). 1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine (DiD) and 3-(4,5-dimethyl-2-tiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were obtained from Beyotime Biotechnology (Beijing, China). 4,6-diamidino-2-phenylindole (DAPI) was purchased from Solarbio Science & Technology (Beijing, China). Low-melting-point agarose was purchased from Gibco (USA). Horseradish peroxidase-conjugated goat anti-rabbit lgG as well as rabbit primary antibodies against GAPDH, paxillin, LC3, p62, MMP2, CD31, α-SMA, and IL-6 were obtained from Abways (Shanghai, China). All other chemicals were of analytical grade.

Chen X., Tao Y., He M., Deng M., Guo R., Sheng Q., Wang X., Ren K., Li T., He X., Zang S., Zhang Z., Li M, & He Q. (2021). Co-delivery of autophagy inhibitor and gemcitabine using a pH-activatable core-shell nanobomb inhibits pancreatic cancer progression and metastasis. Theranostics, 11(18), 8692-8705.

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Synthesis and Purification of Cy5.5-Labeled Lung Targeting Peptides

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Linear Cy5.5 labeled lung targeting peptides (LTPs) were synthesized on a Liberty CEM microwave synthesizer using Fluorenylmethyloxycarbonyl (FMOC) chemistry. Stepwise addition of each FMOC protected amino acid was accomplished on an amide resin as solid support using Ethyl-(2Z)-2-cyano-2-hydroxyiminoacetate/N,N-Diisopropylcarbodiimide (Oxyma/DIC) activation chemistry. The N-terminal amino group of the resin bound LTP peptides were then conjugated with Cy5.5-NHS (Lumiprobe Corporation) in DIPEA/DMF. Final cleavage of Cy5.5-LTP from the resin with Trifluoroacetic acid : Triisopropylsilane : H2O (TFA:TIPS:H2O-90:25:25) was followed by precipitation in Diethyl Ether (EtO2). The resulting crude Cy5.5-LTP peptides were purified by semi-preparative C-5 RP-HPLC on a Waters Delta Prep 4000 chromatography system using standard Acetonitrile/0.1%TFA gradient conditions.
MALDI-TOF analysis of the purified conjugates on an Applied Biosystems Voyager workstation using α-Cyano-4-hydroxycinnamic acid (CHCA) matrix allowed for confirmation of the expected mass and identity of the final product.

McCandless K., Mishra S., Stiltner J., Feldman K.S., Yagi H., Yurko R., Islam K., Brown J.M., Frizzell R, & Zahid M. (2021). Novel Lung Targeting Cell Penetrating Peptides as Vectors for Delivery of Therapeutics. Research Square.

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Exosome Labeling and Cellular Uptake

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Col M-exo was labeled Cyanine5.5-N-hydroxysuccinimide (Cy5.5-NHS) ester (#67020, Lumiprobe, Hunt Valley, MD, USA) regarding the manufacturer’s protocol. The exosomes and Cy5.5-NHS-ester were mixed and incubated overnight. To remove the unattached Cy5.5-NHS-ester, the mixture was centrifuged at 20 psi using Airfuge Air-Driven Ultracentrifuge (340400, Beckman Coulter, Brea, CA, USA) and the supernatant was discarded. This washing step was repeated several times. HDFs were seeded in a 35 mm confocal dish at a density of 2 × 10⁵ cells per dish. The cells were treated with labeled exosomes (0.05, 0.1 mg/mL) and incubated for 1, 4, 16, and 24 h. Then the cells were observed with a confocal microscope (TCS SP8, Leica, Wetzlar, Germany). The fluorescence intensities were evaluated using an image analyzer.

Han G., Kim H., Kim D.E., Ahn Y., Kim J., Jang Y.J., Kim K., Yang Y, & Kim S.H. (2022). The Potential of Bovine Colostrum-Derived Exosomes to Repair Aged and Damaged Skin Cells. Pharmaceutics, 14(2), 307.

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Fluorescent Labeling of Oligonucleotides and Proteins

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Oligonucleotides
(ASO and cODN) modified at the 3′ end with an amino group were
labeled with an N-hydroxy succinimide-modified fluorophore (fluorophore-NHS).
The fluorophore-NHS (either Cy3-NHS or Cy5.5-NHS) (Lumiprobe, cat#
21020 and 47020, respectively) was mixed with the ODN in the presence
of DMSO and 0.1 M HEPES (pH 8) and then incubated overnight at 650
rpm at RT. Excess fluorophore and unconjugated ODN were removed using
ethanol precipitation^{22 (link)} and reversed-phase
high-performance liquid chromatography (RP-HPLC), respectively. The
fluorophore-labeled ODN concentration was determined by measuring
the absorbance at 260 nm with a correction factor (CF₂₆₀) of 0.04 for Cy3-NHS and 0.07 for Cy5.5-NHS.
rHA was labeled
nonspecifically on the amino groups of lysine residues. rHA-cODN conjugate
or the rHA-cODN/ASO biomolecular assembly was mixed with the fluorophore-NHS
in HEPES buffer (pH 8) and incubated overnight at 650 rpm at RT. Excess
fluorophore was then removed by spin filtration through a 10 kDa cutoff
membrane filter (Amicon Ultra, Merck Millipore Ltd., cat# UFC501096).

Elkhashab M., Dilek Y., Foss M., Creemers L.B, & Howard K.A. (2024). A Modular Albumin-Oligonucleotide Biomolecular Assembly for Delivery of Antisense Therapeutics. Molecular Pharmaceutics, 21(2), 491-500.

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Synthesis and Characterization of PLGA Nanoparticles

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Poly(d,l-lactide-co-glycolide) 50:50 was from Evonik Industries (Birmingham, AL, USA). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-ethyldiisopropylamine (N,N-diisopropylethylamine) (DIEA, 99%), thiazolyl blue tetrazolium bromide (MTT, 98%) were from Alfa Aesar (Heysham, England). N-hydroxysuccinimide (NHS, 98%) was from Acros Organics Co. Inc. (Fair Lawn, NJ, USA). Cy5.5-NHS was from Lumiprobe Corporation (Hallandale Bench, FL, USA). T7-FITC peptide (MW 1395.57 g/mol, FITC-Asp-His-Ala-Ile-Tyr-Pro-Arg-His-OH) and R9-peptide (MW 1422.3 g/mol Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-OH) were synthesized by Kelowna International Scientific Inc. (Taipei, Taiwan). Polystyrenes (MW 770, 2430, 3700, 13,700, 18,700, 29,300, 44,000, 114,200 g/mol) were from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA). Poly(vinyl alcohol) (PVA, 88% hydrolyzed) was from Acros Organics Co. Inc. (Fair Lawn, NJ, USA). Phosphotungstic acid (PTA) was from Electron Microscopy Sciences (Hatfield, PA, USA). The bEnd.3 and U87-MG cell lines were from Bioresource Collection and Research Center (Hsinchu, Taiwan). Dulbecco’s Modified Eagle Medium (DMEM) powder was from Thermo Fisher Scientific Inc. (Grand Island, NY, USA).

Lo Y.C, & Lin W.J. (2021). Benefit of a Short Chain Peptide as a Targeting Ligand of Nanocarriers for a Brain-Driven Purpose. Pharmaceutics, 13(8), 1249.

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Tumor-targeted Fluorescent Peptide Imaging

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Five to six-week-old Balb/c female nude mice were subcutaneously administered approximately 1 × 10⁷ SKBR3 cells into the right hind leg, to establish xenografted tumors. Thereafter, tumor size was periodically measured with a caliper, and mice with tumors of 6−8 mm in diameter were selected for the following small animal experiments. Cy5.5–NHS (Lumiprobe) was used to label peptides. Either Cy5.5-peptides or the control Cy5.5 (1 μM, 200 μL) was intravenously injected into tumor-bearing nude mice via the tail vein. The mice were anesthetized and fluorescence signals measured using the small animal in vivo imaging system 30 min postinjection. Three mice were used for each peptide and for control. Near-infrared fluorescence imaging of tumor-bearing nude mice were taken with an exposure time of 50 ms, using the Cy5.5 filter sets (excitation: 673 nm; emission: 707 nm), and the intensities were quantified using the same software. Then, fluorescence images of the main organs and of tumors dissected from nude mice were individually taken as above.

Yang X., Wang Z., Xiang Z., Li D., Hu Z., Cui W., Geng L, & Fang Q. (2017). Peptide probes derived from pertuzumab by molecular dynamics modeling for HER2 positive tumor imaging. PLoS Computational Biology, 13(4), e1005441.

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Fluorescent Labeling of GKRK-APO for In Vitro and In Vivo Analysis

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To monitor the in vitro and in vivo behaviors, GKRK-APO or APO was labeled with the fluorescent dye Cy5.5 through NHS-amino coupling reaction. Briefly, the Cy5.5-NHS (Lumiprobe, Hunt Valley, MD) was dissolved in dry DMSO and added to GKRK-APO or APO solution (PBS, pH 8.0), at a dye to GKRK-APO molar ratio of 10:1. The mixture was gently stirred overnight at 4 °C in the dark and then dialyzed in a dialysis bag to remove free dyes.

Zhai M., Wang Y., Zhang L., Liang M., Fu S., Cui L., Yang M., Gong W., Li Z., Yu L., Xie X., Yang C., Yang Y, & Gao C. (2018). Glioma targeting peptide modified apoferritin nanocage. Drug Delivery, 25(1), 1013-1024.

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