Ki67 alexafluor700

Manufactured by BD

Ki67–AlexaFluor700 is a fluorescently labeled antibody that can be used to detect the Ki67 protein, a marker of cellular proliferation. The AlexaFluor700 dye is used to label the antibody, providing a means for visualization and quantification of Ki67 expression in samples.

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2 protocols using ki67 alexafluor700

Comprehensive Immune Profiling of Pembrolizumab Therapy

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PBMC samples at the indicated visits pre- and post-pembrolizumab treatment were thawed and stained with a fixable Aqua viability dye (Invitrogen) and a cocktail of antibodies to the following surface markers: CD8–Qdot605 (Invitrogen, 3B5), CD4–Qdot655 (Invitrogen, S3.5), PD-1–PE (BD, MIH4), LAG-3–FITC (Enzo, 17B4), ICOS–PE-Cy7 (eBioscience, ISA-3), TIM-3–APC (R&D Systems, 344823). Cells were next fixed and permeabilized with the FOXP3/Ki67 Fixation/Permeabilization Concentrate and Diluent (eBioscience), and subsequently stained intracellularly with CD3–BV570 (Biolegend, UCHT1), Ki67–AlexaFluor700 (BD), FOXP3-eFluor450 (eBioscience), and CTLA-4–PerCP–eFluor710 (eBioscience). Stained cells were acquired on a BD Biosciences LSRFortessa and analysed using FlowJo software (FlowJo, LLC).

Huang A.C., Postow M.A., Orlowski R.J., Mick R., Bengsch B., Manne S., Xu W., Harmon S., Giles J.R., Wenz B., Adamow M., Kuk D., Panageas K.S., Carrera C., Wong P., Quagliarello F., Wubbenhorst B., D’Andrea K., Pauken K.E., Herati R.S., Staupe R.P., Schenkel J.M., McGettigan S., Kothari S., George S.M., Vonderheide R.H., Amaravadi R.K., Karakousis G.C., Schuchter L.M., Xu X., Nathanson K.L., Wolchok J.D., Gangadhar T.C, & Wherry E.J. (2017). T-cell invigoration to tumour burden ratio associated with anti-PD-1 response. Nature, 545(7652), 60-65.

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Comprehensive Immune Cell Profiling

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GICs and PBMCs were stained for CD3-PE-Cy5.5 (eBioscience), CD4-BV711, CD8-PerCP, CD25-BV605, CD45RA-BV570, CCR7-BV650, CD39-BV421, PD-1-APC-Cy, BTLA-PE, Tim-3-PE-Cy7 (all Biolegend), LAG3 (Enzo Life Sciences, Lörrach, Germany), CTLA4-PE-CF594 (BD) and Live/dead-Aqua dye (Life technologies, Carlsbad, CA) or with isotype controls. Cells were fixed and permeabilized, followed by ICS using Foxp3-FITC (eBioscience) and Ki67-Alexa Fluor 700 (BD) and measured on an LSR Fortessa (BD).

Rammensee H.G., Wiesmüller K.H., Chandran P.A., Zelba H., Rusch E., Gouttefangeas C., Kowalewski D.J., Di Marco M., Haen S.P., Walz J.S., Gloria Y.C., Bödder J., Schertel J.M., Tunger A., Müller L., Kießler M., Wehner R., Schmitz M., Jakobi M., Schneiderhan-Marra N., Klein R., Laske K., Artzner K., Backert L., Schuster H., Schwenck J., Weber A.N., Pichler B.J., Kneilling M., la Fougère C., Forchhammer S., Metzler G., Bauer J., Weide B., Schippert W., Stevanović S, & Löffler M.W. (2019). A new synthetic toll-like receptor 1/2 ligand is an efficient adjuvant for peptide vaccination in a human volunteer. Journal for Immunotherapy of Cancer, 7, 307.

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