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8 protocols using cy2 and cy5 conjugated antibodies

1

Tau Protein Characterization in AD Brain

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DMEM was purchased from Mediatech, and FBS was obtained from Atlas Biologicals. Antibiotic-antimycotic was purchased from MilliporeSigma. Recombinant human Tau-441 was purchased from AnaSpec. Primary antibodies used in the study are listed in Supplemental Table 1. Cy2- and Cy5-conjugated antibodies and fluorophore-tagged secondary antibodies were obtained from Jackson ImmunoResearch Laboratories Inc. AD brain–derived insoluble tau was provided by Virginia M.-Y. Lee, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
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2

Immunofluorescence Staining of Dopaminergic Neurons

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Roswell Park Memorial Institute (RPMI) medium were purchased from Mediatech (Washington, DC). Fetal bovine serum (FBS) was obtained from Atlas Biologicals (Fort Collins, CO). Antibiotic-antimycotic and MPTP were purchased from Sigma-Aldrich (St. Louis, MO). Recombinant mouse RANTES was purchased from R&D Systems (Minneapolis, MN). Anti-CD4 antibody was purchased from eBioscience. Rabbit anti-tyrosine hydroxylase (TH) antibody was purchased from Pel-Freeze biologicals (Rogers, AR). Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno-Research Laboratories (West Grove, PA).
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3

Quantification of Chemokines and Immune Markers

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Mouse RANTES and eotaxin ELISA kits as well as mouse recombinant RANTES and eotaxin were purchased from R&D Systems (Minneapolis, MN). Anti-CD4 antibodies were purchased from eBioscience. Rabbit anti-TH antibody was purchased from Millipore. Anti-Iba-1 antibody was purchased from Abcam. Anti-GFAP antibody was purchased from DAKO. Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno Research Laboratories (West Grove, PA).
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4

Antibody Characterization for Mitochondrial Proteins

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Rabbit polyclonal anti-PGC1α, mouse monoclonal anti-Nrf1 and rabbit polyclonal anti-TFAM antibodies were obtained from Abcam (Cambridge, MA). Anti-tyrosine hydroxylase (TH), anti-p300 and anti-NeuN antibodies were obtained Millipore (Billerica, MA). Rabbit monoclonal antibodies against phospho-CREB, total CREB and CREB-binding protein (CBP) were purchased from Cell Signaling (Beverly, MA). Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno Research Laboratories (West Grove, PA).
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5

Immunohistochemical Analysis of Neuroinflammation

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Phosphate buffer saline (PBS), trypsin, DMEM/F-12, and antibodies against CD68 were purchased from Thermo Fisher Scientific (Waltham, MA). Fetal bovine serum (FBS) was bought from Atlas Biologicals (Fort Collins, CO). Antibiotic-antimycotic was purchased from Sigma (St. Louis, MO). Recombinant human α-syn was purchased from Anaspec (Fremont, CA). Antibodies against anti-goat Iba1 were purchased from Abcam (Waltham, MA). Dako antibodies against glial fibrillary acidic protein (GFAP) were bought from Agilent (Santa Clara, CA). Antibodies against myelin basic protein (MBP) and microtubule associated protein 2 (MAP2) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno-Research Laboratories (West Grove, PA).
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6

Immunodetection of Neurodegeneration Markers

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Rabbit anti-mouse iNOS antibodies were obtained from Calbiochem, Gibbstown, NJ. Goat anti-glial fibrillary acidic protein (GFAP) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-mouse Parkin and goat anti-mouse DJ-1 were purchased from Abcam (Cambridge, MA). Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno Research Laboratories (West Grove, PA).
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7

Immunodetection of Immune Markers

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Human chemokine array kit (Enzyme-Linked ImmunoSpot; ELISPOT) was purchased from Raybiotech. Human chemokine ELISA array kit was purchased from SAbiosciences. Anti-CD3, CD4 and CD8 antibodies were purchased from e biosciences. Rabbit anti-TH antibody was purchased from Calbiochem (a Millipore Company). Anti-Iba-1 antibody was purchased from Abcam. Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno Research Laboratories (West Grove, PA).
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8

Immunostaining of alpha-synuclein in cell culture

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Dulbecco’s Modified Eagle Medium (DMEM) medium was purchased from Mediatech (Washington, DC) and fetal bovine serum (FBS) was obtained from Atlas Biologicals (Fort Collins, CO). Antibiotic-antimycotic was purchased from Sigma-Aldrich (St. Louis, MO). Recombinant human α-syn was purchased from Anaspec (Fremont, CA). Rabbit anti-tyrosine hydroxylase (TH) antibody was purchased from Pel-Freeze biologicals (Rogers, AR) and mouse anti-TH antibody was procured from Immunostar (Hudson, WI). Mouse α-syn antibody was purchased from BD Bioscience (San Jose, CA) (Supplementary Table S1). Cy2- and Cy5-conjugated antibodies were obtained from Jackson Immuno-Research Laboratories (West Grove, PA).
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