Pacificblue nhs ester

Manufactured by Thermo Fisher Scientific

The PacificBlue-NHS Ester is a fluorescent labeling reagent. It is designed for covalent attachment to primary amines on proteins, peptides, and other biomolecules. The Pacific Blue fluorophore has an excitation maximum of 410 nm and an emission maximum of 455 nm.

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Lab products found in correlation

Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (3 mentions) Pacific orange nhs ester, by Thermo Fisher Scientific (3 mentions) Alexafluor af 488 nhs ester, by Thermo Fisher Scientific (3 mentions) Live dead fixable violet dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Angiosense 750, by PerkinElmer (1 mentions)

Close spelling variants of this product

NHS-esters (2 citations)

4 protocols using pacificblue nhs ester

Multiparametric Immunophenotyping of Cells

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Cells were live/dead stained using the Live/Dead Fixable Near IR Dead
Cell Stain Kit (Life Technologies). For surface stains, cells were stained for
30 minutes on ice with indicated antibodies. Clones, sources & dilutions
can be found in the Supplementary Table 5. For intracellular stains, samples were fixed
at the indicated time after stimulation in 2% paraformaldehyde, surface
stained with CD25-biotin and CD4-BUV395, fixed again and permeabilized with ice
cold 90% methanol at −20 °C overnight. Samples were then
barcoded using Pacific Orange-NHS ester (0.33 or 5 μg/mL), Pacific
Blue-NHS Ester (0.67 or 10 μg/mL), and AlexaFluor (AF) 488-NHS Ester
(0.26 or 2 μg/mL) (Life Technologies), as previously described³⁶. Intracellular antigens were
then stained for 30 minutes at 23 °C with antibodies indicated in Supplementary Table 5.
Samples were acquired on a BD LSR Fortessa and analyzed in FlowJo (Tree
Star).

Smith G.A., Uchida K., Weiss A, & Taunton J. (2016). Essential biphasic role for JAK3 catalytic activity in IL-2 receptor signaling. Nature chemical biology, 12(5), 373-379.

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Multiparametric Immunophenotyping of Cells

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Smith G.A., Uchida K., Weiss A, & Taunton J. (2016). Essential biphasic role for JAK3 catalytic activity in IL-2 receptor signaling. Nature chemical biology, 12(5), 373-379.

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Multiparametric Flow Cytometry Analysis

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Cells were live/dead-stained using the Live/Dead Fixable Near IR Dead Cell Stain Kit or Live/Dead Fixable Violet Dead Cell Stain Kit (Life Technologies). For surface stains, cells were stained for 30 minutes on ice with the indicated antibodies. Information on antibody clones, sources, and working dilutions are listed in Table S1. For intracellular stains, samples were fixed at the indicated time after stimulation in 2% paraformaldehyde, surface stained with CD8a-BUV737 and CD4-BUV395, fixed again, then permeabilized with ice cold 90% methanol at −20 °C overnight. Samples were then barcoded using Pacific Orange-NHS ester (0.33 or 5 μg/mL), Pacific Blue-NHS Ester (0.67 or 10 μg/mL), and AlexaFluor (AF) 488-NHS Ester (0.26 or 2 μg/mL) (Life Technologies), as previously described(5 (link)). Intracellular antigens were then stained for 30 minutes at 23 °C with antibodies indicated in Table S1. Samples were acquired on a BD LSR Fortessa SORP and analyzed in FlowJo (Tree Star).

Smith G.A., Taunton J, & Weiss A. (2017). IL-2Rβ Abundance Differentially Tunes IL-2 Signaling Dynamics in CD4+ and CD8+ T Cells. Science signaling, 10(510), eaan4931.

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Intravital Imaging of Tumor Vasculature

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Xenografts were imaged as previously described (40 ), and all animal experiments were carried out in accordance with guidelines from the Institutional Subcommittee on Research Animal Care. Briefly, dorsal skin window chambers were surgically implanted in 6–8 week old nude mice. Approximately 3–4 million cells were suspended in a 50% Matrigel and 50% PBS solution and injected under the fascia and allowed to grow for 1–2 weeks. When tumors were vascularized and reached 1–2 mm in size, mice were anesthetized using 1–2% isoflurane in 2 L/min oxygen and a tail vein catheter was inserted. The mouse was moved to a heated microscope stage where the vital signs were continuously monitored. Angiosense-750 (Perkin Elmer) or 250 μg of 500 kDa amino-dextran labeled with Pacific Blue NHS ester (Life Technologies) was injected via tail vein and used to visualize the vasculature. Once the region of interest was selected, a time-lapse imaging sequence was initiated, followed by tail vein injection of the probe. 75 nmol of probe was formulated in 30 μL of 1:1 dimethylacetamide:solutol solution and 112.5 μL of phosphate buffered saline and sonicated.

Thurber G.M., Reiner T., Yang K.S., Kohler R, & Weissleder R. (2014). Effect of Small Molecule Modification on Single Cell Pharmacokinetics of PARP Inhibitors. Molecular cancer therapeutics, 13(4), 986-995.

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