From HEK293T lysates, SEC fraction 9 from PS19 mice, and AD and PSP human samples, tau was immunoprecipitated by using 2 µg of biotinylated HT7 antibody (Thermo Fisher, MN1000b), for every 100 ng of human tau quantified by ELISA. IgG isotype control antibodies (BioLegend, 400104) were used for comparison. The IP and flow-through samples were then subjected to further analysis.
Igg isotype control antibody
Manufactured by BioLegend
Sourced in United States
IgG isotype control antibodies are laboratory reagents used to establish baseline or non-specific binding in flow cytometry and other immunoassay experiments. These antibodies are of the same immunoglobulin class (IgG) as the target-specific antibody, but they do not bind to the antigen of interest.
Automatically generated - may contain errors
Lab products found in correlation
Ht7 antibody, by Thermo Fisher Scientific (1 mentions)
Fitc conjugated anti human cd14 antibody, by BioLegend (1 mentions)
Facsarray, by BD (1 mentions)
Alexa fluor 647 conjugated anti human cd45 antibody, by BioLegend (1 mentions)
Pe conjugated anti human cd163 antibody, by BioLegend (1 mentions)
Goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
Mg6 cells, by RIKEN BioResource Center (1 mentions)
Pkh26 red fluorescent cell linker kit, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Recombinant rat beta ngf, by R&D Systems (1 mentions)
6 protocols using igg isotype control antibody
1
Tau Immunoprecipitation from Diverse Samples
2
Multicolor Flow Cytometry of Immune Cells
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The samples were washed with PBS containing 2% FBS and centrifuged at 300 × g, 4°C for 5 minutes. Dead cells were stained with APC-Cy7-conjugated Zombie NIR (BioLegend, #423105), and the cells were subsequently stained with the following surface markers for 30 minutes at 4°C: Alexa Fluor 647-conjugated anti-human CD45 antibody (BioLegend), FITC-conjugated anti-human CD14 antibody (BioLegend, #301803, RRID:AB_314185), and PE-conjugated anti-human CD163 antibody (BioLegend). As the controls, IgG-isotype control antibodies (BioLegend, #400112) were used. Cells were analyzed by flow cytometry (FACS Array, Becton Dickinson), and data were reanalyzed with Flow Jo software (Tree Star).
3
CD4+ T Cell Depletion Model
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Experimental animals were depleted of CD4+ cells using a rat anti-mouse CD4 monoclonal IgG2b antibody (clone GK1.5; BioLegend, San Diego, CA, USA). The antibody was administered i.p. (300 μg per mouse) 24 h before ConA administration. Control animals were treated with IgG isotype control antibodies (BioLegend).
4
TIM3 Pathway Modulation in THP-1 Cells
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The TIM3 pathway was interfered with by TIM3 blocking antibody, with the THP-1 cells being incubated with 10 μg/mL of TIM3 blocking antibody (#345004, BioLegend, San Diego, CA, USA) or IgG isotype control antibody (#400153, BioLegend, San Diego, CA, USA) for 24 h. Then, the CLEC7A, CCL13, and IL-6 mRNA levels were measured by RT-qPCR or ELISA.
5
SLAMF5 Stimulation of Dendritic Cells
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For SLAMF5 stimulation moDCs adjusted to 107 ml−1 were suspended in complete RPMI-1640 medium containing 10 µg ml−1 anti-SLAMF5 antibody (clone 152-1D5; LifeSpan BioSciences, Cat. No. LS-C134663) or an IgG isotype control antibody (BioLegend, Cat. No. 400124) at 4°C for 45 min. Thereafter, following thorough washing procedures, cells were reseeded into 24-well cell culture plates and incubated in complete medium in the presence of 10 µg ml−1 F(ab′)2 of goat anti-mouse IgG (Jackson ImmunoResearch Laboratories, Cat. No. 115-006-062) at 37°C for 2 h.
6
Characterization of Phagocytic Cell Lines
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PC12 cells and MG6 cells27 (link)28 (link) were obtained from RIKEN BRC, Japan. PC12 cells were maintained in DMEM (Nakarai, Japan) supplemented with 10% FBS (Biowest) and 10% horse serum (Life technologies), and MG6 cells were maintained in DMEM supplemented with 10% FBS, 10 μg/ml insulin and 100 μM 2-mercaptoethanol. Recombinant rat beta-NGF and pan-caspase OPH inhibitor Q-VD26 (link) were from R&D systems. Monoclonal antibody against C3a/C3 (clone K13/16) and IgG isotype control antibody were from BioLegend. The NIH3T3 transformants expressing Tim4 (3T3/Tim4) and rD89E were described previously31 (link)32 (link). When indicated, membranes of MG6 cells or exosomes were labelled with PKH26 red fluorescent cell linker kit (Sigma) according to the manufacturer's instructions.
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