Table 2 Immunohistochemistry panels for analysis of microglial changes in the human ALS and hTDP-43-ΔNLS ‘rNLS’ mouse brains
PANEL 1 Human multiplexed quantification | |||
|---|---|---|---|
Round 1 primary antibody | Round 1 secondary antibody | Round 2 primary antibody | Round 2 secondary antibody |
Mouse IgG2b anti-L-ferritin (1:1,000; Abcam, Ab218400) | Goat anti-mouse IgG2b AlexaFluor® 488 (1:500; ThermoFisher, A21141) | Mouse IgG1 anti-TDP-43, phospho Ser409/410 (1:1,000; Cosmobio, CAC-TIP-PTD-M01) | Goat anti-mouse IgG1 AlexaFluor® 488 (1:500; ThermoFisher, A21121) |
Mouse IgG1 anti-HLA-DQ, DP, DR (1:500; DAKO, M0775) | Goat anti-mouse IgG1 AlexaFluor® 546 (1:500; ThermoFisher, A21123) | Guinea pig anti-NeuN (1:500; Sigma Aldrich, ABN90) | Goat anti-guinea pig AlexaFluor® 546 (1:500; ThermoFisher, A11074) |
Mouse IgG3 anti-CD68 (1:200; Abcam, Ab783) | Goat anti-mouse IgG3 AlexaFluor® 594 (1:500; ThermoFisher, A21155) | Rabbit anti-GFAP (1:2,000; DAKO, Z0334) | Goat anti-rabbit AlexaFluor® 594 (1:500; Jackson Laboratories, 111–586-144) |
Rabbit anti-CD74 (1:1,000; Abcam, Ab64772) | Goat anti-rabbit AlexaFluor® 647 (1:500; Jackson Laboratories, 111–605-144) | UEA-1 Lectin, biotinylate (1:1,000; Sigma-Aldrich, L8262) | Streptavidin AlexaFluor® 647 (1:500; Jackson Laboratories, 016–170-084) |
Chicken anti-Iba1 (1:1,000; Synaptic Systems, 234,009) | Goat anti-chicken AlexaFluor® 800-plus (1:500; ThermoFisher, A32935) |  | |
PANEL 2 Human confocal | |
|---|---|
Primary antibody | Secondary antibody |
Mouse IgG2b anti-L-ferritin (1:1,000; Abcam, Ab218400) | Goat anti-mouse IgG2b AlexaFluor® 488 (1:500; ThermoFisher, A21141) |
Mouse IgG3 anti-CD68 (1:200; Abcam, Ab783) | Goat anti-mouse IgG3 AlexaFluor® 594 (1:500; ThermoFisher, A21155) |
Chicken anti-Iba1 (1:1,000; Synaptic Systems, 234,009) | Goat anti-chicken IgG AlexaFluor® 647 (1:500; ThermoFisher, A21449) |
PANEL 3 Mouse multiplexed quantification | |||
|---|---|---|---|
Round 1 primary antibody | Round 1 secondary antibody | Round 2 primary antibody | Round 2 secondary antibody |
Rabbit anti-CD68 (1:1,000; Abcam, Ab215212) | Donkey anti-rabbit IgG AlexaFluor® 488 (1:500; Jackson Laboratories, 711–545-152) | Rabbit anti-pTDP-43, Ser 409/410 (1:3,000; Cosmo Bio, TIP-PTD-P02) | Donkey anti-rabbit AlexaFluor® 488 (1:500; Jackson Laboratories, 711–545-152) |
Goat anti-L-ferritin (1:1,000; Lifespan Biosciences, LS-B4383-50) | Donkey anti-goat AlexaFluor® 546 (1:500; ThermoFisher, A11056) | – | – |
Guinea pig anti-NeuN (1:500; Sigma Aldrich, ABN90) | Donkey anti-guinea pig AlexaFluor® 594 (1:500; Jackson Laboratories, 706–585-148) | – | – |
Chicken anti-Iba1 (1:1,000; Synaptic Systems, 234,009) | Donkey anti-chicken AlexaFluor® 647 (1:500; Jackson Laboratories, 703–605-155) | Goat anti-Iba1 (1:1,000; Abcam, Ab5076) | Donkey anti-goat AlexaFluor® 647 (1:500; Jackson Laboratories, 705–605-947) |