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Biotinylated antibodies

Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom

Biotinylated antibodies are antibodies that have been chemically modified by the addition of biotin, a small molecule that binds strongly to the protein avidin. This allows the antibodies to be easily detected or captured in various immunoassay and affinity-based applications.

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3 protocols using biotinylated antibodies

1

Mitochondrial Dynamics in Cell Viability Analysis

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Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), L-glutamine (Q), and penicillin/streptomycin (P/S) were obtained from Invitrogen (Carlsbad, CA). MitoTracker Green FM and MitoTracker Red CMXRos, were purchased from Molecular Probes (Eugene, OR). Manganese Chloride (MnCl2) was obtained from Sigma-Aldrich (St. Louis, MO). The CellTiter® 961 AQueous Non-Radioactive Cell Proliferation Assay kit and CellTiter® Glo Luminescent Cell Viability Assay kit were obtained from Promega (Madison, WI). The CD11b magnetic separation kit was purchased from Stem Cell Technologies (Vancouver, Canada). Tyrosine hydroxylase (TH) (AB_2201528) and glial fibrillary acidic protein (GFAP) (AB_2109815) antibodies were purchased from EMD Millipore. Inducible nitric oxide synthase (iNOS) (AB_631831) antibody was purchased from Santa Cruz Biotechnologies (Dallas, TX). Mitofusin-2 (Mfn2) (#11925) was purchased from Cell Signaling Technologies. All the standards used for the Luminex multiplex cytokine assay were purchased from PeproTech Inc (Rocky Hill, NJ). Streptavidin-Biotin and biotinylated antibodies used for Luminex were purchased from eBioSciences (San Diego, CA). MA was obtained from Dr. Kalyanaraman (Medical College of Wisconsin, Milwaukee).
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2

Multicolor Immunohistochemistry of Pancreatic Immune Cells

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Pancreata embedded in OCT compound (Cellpath, Newtown, UK) were frozen in isopentane (Sigma) cooled with liquid nitrogen. 8–10 μm sections were fixed in acetone before staining with biotinylated antibodies to CD4, CD8, CD11b, CD11c, GR-1 and B220 (all from eBioscience, Altrincham, UK). Staining was detected using ABC reagent and DAB solution from Vector Labs (Peterborough, UK). Images acquired on a Zeiss Axiovert A1 microscope using Zen software. No cropping or alterations of the images other than changing contrast was used.
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3

Multimodal Characterization of Macrophage Response

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Dulbecco’s modified Eagle’s medium (DMEM), DMEM-F12, fetal bovine serum (FBS), L-glutamine (Q), penicillin/streptomycin (P/S), Sodium Pyruvate (SP), and Non-Essential Amino Acids (NEAA) were obtained from Invitrogen (Carlsbad, CA). CellTiter Glo Luminescent Cell Viability Assay kit was obtained from Promega (Madison, WI). The CD11b magnetic separation kit was purchased from Stem Cell Technologies (Vancouver, Canada). All standards used for the Luminex multiplex cytokine assay were purchased from PeproTech, Inc (Rocky Hill, NJ). Streptavidin-Biotin and biotinylated antibodies used for Luminex were purchased from eBioSciences (San Diego, CA). Phagocytosis assay kit was purchased from Cayman Chemicals (Ann Arbor, MI).
For qRT-PCR, the QuantStudio3 Real Time PCR system from Thermo Fisher (Waltham, MA) was used. For MTS and Griess assays, a Molecular Devices (Sunnyvale, CA) plate reader was used. For scanning Western blot, a Li-COR Odyssey gel imager (Lincoln, NE) was used. Cell culture supplies were purchased from Corning (Corning, NY). For immunocytochemistry studies, the images were obtained using a Nikon (Model: TE-2000U) inverted fluorescent microscope and images were obtained using a Spot Digital Camera (Sterling Heights, MI).
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