recommendations antibody polyclonal goat igg anti rat tnfa  (R&D Systems)

Journal: eLife

Article Title: Microglia TREM2R47H Alzheimer-linked variant enhances excitatory transmission and reduces LTP via increased TNF-α levels

doi: 10.7554/elife.57513

Figure Lengend Snippet: Figure 7. LTP is impaired in RH mutant rats, and the impairment could be rescued by application of anti-TNF-a antibody. (A) The input-output slope is significantly increased in Trem2R47H/R47H rats [two-way ANOVA, stimulation intensity x genotype interaction F(6, 144)=6.745, p<0.0001****; post-hoc Sidak’s multiple comparisons test: 1.2 mV p=0.0175*; 1.4 mV: p=0.0021**; 1.6 mV: p=0.0001***] Representative traces of fEPSPs in response to increasing stimulus from 0.4 to 1.6 mA are shown on the top. (B) LTP is impaired in Trem2R47H/R47H rats. Average traces of the baseline and the last 5mins of LTP are shown on top. (C) Plot of fEPSP slope change of the last 10 min of LTP in B (unpaired t test, p<0.0001****). (D) The increase of input- out slope in Trem2R47H/R47H rats is reversed by application of anti-TNF-a [ANOVA for repeated measures F(18, 300)=6.579, p<0.0001****; post-hoc Tukey’s multiple comparisons test: 0.8 mV RH/RH + Isotype vs. w/w + anti-TNF-a p=0.0479*; 1.0 mV RH/RH + Isotype vs. RH/RH + anti-TNF-a p=0.0061**, RH/RH + Isotype vs. w/w + anti-TNFa p=0.0072**, RH/RH + Isotype vs. w/w + Isotype p=0.0113*; 1.2 mV RH/RH + Isotype vs. RH/RH + anti-TNFa p=0.0009***, RH/RH + Isotype vs. w/w + anti-TNF-a p=0.0009***, RH/RH + Isotype vs. w/w + Isotype p=0.0029**; 1.4 mV RH/RH + Isotype vs. RH/RH + anti-TNF-a p<0.0001****, RH/RH + Isotype vs. w/w + anti-TNF-a p<0.0001****, RH/RH + Isotype vs. w/w + Isotype p=P < 0.0001****; 1.6 mV RH/RH + Isotype vs. RH/RH + anti-TNF-a p<0.0001****, RH/RH + Isotype vs. w/w + anti-TNF-a P p<0.0001****, RH/RH + Isotype vs. w/w + Isotype p<0.0001****]. Representative fEPSP traces are shown on top. (E) The impaired LTP is restored by application of anti-TNF-a antibody. The average traces of the baseline and the last 5mins of LTP are shown on top. (F) Plot of fEPSP slope change of the last 10 min of LTP in E [one-way ANOVA, F(3,

Article Snippet: Aging Cell 18: e13033 Rat App allele with humanize Ab region Genetic reagent (Rattus Norvegicus) Trem2R47H Tambini and D’Adamio, 2020 Sci Rep 10: 4122 Rat Trem2 allele with R47H mutation Commercial assay or kit V-PLEX Plus Ab Peptide Panel 1 Meso Scale Discovery Cat# K15200G Used following manufacturer’s recommendations Commercial assay or kit V-PLEX Proinflammatory Panel 2 Meso Scale Discovery Cat# K15059D Used following manufacturer’s recommendations Commercial assay or kit CD11b/c (Microglia) Micro-Beads, rat antibody Cat# 130-105-634 Miltenyi Biotec RRID:AB_2783886 Used following manufacturer’s recommendations Commercial assay or kit Adult Brain Dissociation Kit Miltenyi Biotec Cat# 130-107-677 Used following manufacturer’s recommendations Commercial assay or kit RNeasy RNA Isolation kit Qiagen Cat# 74106 Used following manufacturer’s recommendations Commercial assay or kit High-Capacity cDNA RT kit Thermo Cat# 4368814) Used following manufacturer’s recommendations Commercial assay or kit TaqMan Fast Advanced Mix Thermo Cat# 4444556 Used following manufacturer’s recommendations Commercial assay or kit Gapdh RealTime PCR Thermo Rn01775763_g1 Used following manufacturer’s recommendations Commercial assay or kit Treml1 RealTime PCR Thermo Rn01511908_g1 Used following manufacturer’s recommendations Antibody Polyclonal Goat IgG anti-Rat TNFa Cat# AF-510-NA R and D Systems RRID:AB_354511 10 ng/ml in ACSF Antibody Polyclonal Goat IgG. antibody Cat# AB-108-C R and D Systems RRID:AB_354267 10 ng/ml in ACSF Software, algorithm LinRegPCR software hartfaalcentrum.nl Software, algorithm pCLAMP10 software Molecular Devices, Software, algorithm Image Lab software Biorad RRID:SCR_014210 Software, algorithm GraphPad Prism RRID:SCR_002798 Ren et al. eLife 2020;9:e57513.

Techniques: Mutagenesis