Cyanine3 (NHS ester), Cyanine3 (maleimide), Cyanine5 (NHS ester) and Cyanine5 (maleimide) were purchased from Lumiprobe. Puromycin was purchased from Cayman Chemical. UltraPure™ 1M Tris–HCI pH 7.5 was obtained from Invitrogen. Other common materials and reagents were purchased from Sigma or Amresco.
Cyanine 5
Manufactured by Lumiprobe
Sourced in Germany, United States
Cyanine 5.5 is a fluorescent dye used in various laboratory applications. It is a member of the cyanine dye family and exhibits a long absorption and emission wavelength range. Cyanine 5.5 can be utilized in fluorescence-based techniques, such as flow cytometry, immunoassays, and nucleic acid detection.
Automatically generated - may contain errors
Lab products found in correlation
Living image software, by PerkinElmer (1 mentions)
Cyanine 5.5 amine, by Lumiprobe (1 mentions)
Ivis lumina series 3, by PerkinElmer (1 mentions)
Ab18184, by Abcam (1 mentions)
Slide a lyzer, by Thermo Fisher Scientific (1 mentions)
Puromycin, by Cayman Chemical (1 mentions)
Ultrapure 1 m tris hci ph 7, by Thermo Fisher Scientific (1 mentions)
Cyanine3 (cy3), by Lumiprobe (1 mentions)
6 protocols using cyanine 5
1
Fluorescent Dyes for Biomolecular Labeling
2
Biodistribution of TPPOH Treatments
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To determine the biodistribution of TPPOH treatments, free TPPOH and TPPOH-X SNPs were labeled with Cyanine 5.5 (Lumiprobe, Hannover, Germany) to allow tracking because the TPPOH emission spectrum overlaps with that of blood. HT-29 tumor-bearing mice, established as described above, were randomly divided into two groups (n = 3) and intravenously injected with 1/100e LD50 for both group: Cy5.5-free TPPOH (3.26 mg/kg) and Cy5.5-TPPOH-X SNPs (1.16 mg/kg TPPOH and 334 mg/kg SNPs). Then 24 h post-injection, the biodistribution was determined using IVIS Lumina quantitative fluorescence imaging system (PerkinElmer, Villepinte, France). Subsequently, the mice were sacrificed and ex vivo biodistribution images of the tumors and major organs were immediately taken. Relative signal intensity in tumors and organs was calculated, using Living Image software (PerkinElmer), as radiant efficiency ([photons/s/cm2/sr]/[μW/cm2]) per pixel of the region of interest (ROI), which was drawn around the respective organ.
3
Synthesis of Cyanine5.5-Mannose Conjugate
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Example 10
N-(Cyanine5.5)-4′-(α-D-mannopyranosyl)-3′-methyl-[1,1′biphenyl]-3-carboxamide chloride (10) 23 was made following the coupling procedure for 7, utilizing 4′-(α-D-mannopyranosyloxy)-3′-methyl-[1,1′biphenyl]-3-carboxylic acid (loc. cit.) (12.4 mg, 0.032 mmol) and HATU (12.2 mg, 0.032 mmol) in dry DMF (2 mL), followed by commercially available Cyanine5.5amine (Lumiprobe, #7000) (20 mg, 0.027 mmol) and DIPEA (28 μL, 0.16 mmol), to give the product in 69% yield.
Formula: C66H77N4O8+ Exact Mass: 1053.57 Molecular Weight: 1054.34
ESI-MS [M]+ calcd for C66H77N4O8+ 1053.57 found 1053.9.
4
Meningioma Tumor Imaging and Targeting
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The growth of meningioma (CH157-MN-FLuc) tumors in xenografted NSG mice was monitored by measuring bioluminescent signal (FLuc) using an IVIS Lumina Series III (PerkinElmer, Waltham, MA, USA) every two days post cells injection. To monitor meningioma targeting, the anti-SSTR2 mAb or ADC was labeled with Cyanine 5.5 (Lumiprobe, Hunt Valley, MD, USA) according to the manufacturer protocol. The Cy5.5-labeled mAb or ADC was intravenously (i.v.) injected into mice via tail vein. At 24 h post injection, the xenograft mice were imaged under IVIS with a wavelength of 660/710 nm (excitation/emission) and an exposure time of 10 s to analyze the meningioma targeting and biodistribution in vivo. The important organs, including brain, heart, lung, kidney, and spleen, were also extracted to collect ex vivo images to check the possible off-target binding.
5
Antibody and Lectin Microarray Preparation
6
Quantification of Primary Amines on CeO2 NPs
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The primary amine groups of CeO2@P2 and CeO2@P3 were quantified by spectrofluorometry (Supplementary Information S4). CeO2@P2 and CeO2@P3 were labelled with Cyanine 5 (Lumiprobe, #23320) in PBS buffer adjusted to pH 8 (2 hours at room temperature while stirring continuously). Particles were washed five times with PBS on an exclusion column (30 kDa) to remove the free Cyanine. The same experimental procedure was carried out with the amine-free particle CeO2@P1 as a control, to validate the washing procedure.
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