Incubation:Article Title: Critical Role of GFR?1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Slides were removed from the − 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFRα1 (1;200;R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- α6 (1:500; BD Biosciences), rat anti-NCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
Article Title: Critical Role of GFR?1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Immunohistochemistry Slides were removed from the − 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFRα1 (1;200;R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- α6 (1:500; BD Biosciences), rat anti-NCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
Article Title: Critical Role of GFR 1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Slides were removed from the 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFR 1 (1;200; R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- 6 (1:500; BD Biosciences), rat antiNCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
Blocking Assay:Article Title: Critical Role of GFR?1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Slides were removed from the − 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFRα1 (1;200;R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- α6 (1:500; BD Biosciences), rat anti-NCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
Article Title: Critical Role of GFR?1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Immunohistochemistry Slides were removed from the − 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFRα1 (1;200;R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- α6 (1:500; BD Biosciences), rat anti-NCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
Article Title: Critical Role of GFR 1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Slides were removed from the 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFR 1 (1;200; R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- 6 (1:500; BD Biosciences), rat antiNCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
Immunofluorescence:Article Title: Noradrenergic innervation across brain regions is altered by aging and by disease progression in a mouse model of Alzheimer's disease neuropathology.
Article Snippet: For immunohistochemical staining using 3,3-diaminobenzidine (DAB) as the chromogen, antibodies included: mouse anti-norepinephrine transporter primary (1:250; monoclonal; catalog # MA5–24647, Invitrogen), mouse anti-tyrosine PLOS One | https://doi.org/10.1371/journal.pone.0340611 February 18, 2026 4 / 17 hydroxylase primary (1:5000; monoclonal; catalog # 22941, ImmunoStar), rabbit anti-amyloid-β primary (1:1000; polyclonal; catalog # 71–5800, Invitrogen), donkey anti-mouse secondary (1:500; polyclonal; catalog # 715-065-150, Jackson ImmunoResearch), and goat anti-rabbit secondary (1:200; clonal; catalog # 31820, Vector Laboratories), with enhanced signal amplification using Avidin-Biotin Complex (Vector Laboratories). .. For immunofluorescence, primary antibodies included: mouse anti-norepinephrine transporter (1:300; catalog # MA5–24647, Invitrogen), mouse anti-tyrosine hydroxylase (1:2000; catalog # 22941, ImmunoStar), rabbit anti-GAP-43 (1:1000; catalog # NB300–143, Novus Biologicals), and rabbit anti-amyloid-β (1:300; catalog # 71–5800, Invitrogen). .. Secondary antibodies included: goat anti-mouse AlexaFluor 488 secondary (1:500; catalog # A-11029, Invitrogen) and goat anti-rabbit AlexaFluor 594 secondary (1:500; catalog # A-11037, Invitrogen).
Article Title: Noradrenergic innervation across brain regions is altered by aging and by disease progression in a mouse model of Alzheimer’s disease neuropathology
Article Snippet: For immunohistochemical staining using 3,3-diaminobenzidine (DAB) as the chromogen, antibodies included: mouse anti-norepinephrine transporter primary (1:250; monoclonal; catalog # MA5–24647, Invitrogen), mouse anti-tyrosine hydroxylase primary (1:5000; monoclonal; catalog # 22941, ImmunoStar), rabbit anti-amyloid-β primary (1:1000; polyclonal; catalog # 71–5800, Invitrogen), donkey anti-mouse secondary (1:500; polyclonal; catalog # 715-065-150, Jackson ImmunoResearch), and goat anti-rabbit secondary (1:200; clonal; catalog # 31820, Vector Laboratories), with enhanced signal amplification using Avidin-Biotin Complex (Vector Laboratories). .. For immunofluorescence, primary antibodies included: mouse anti-norepinephrine transporter (1:300; catalog # MA5–24647, Invitrogen), mouse anti-tyrosine hydroxylase (1:2000; catalog # 22941, ImmunoStar), rabbit anti-GAP-43 (1:1000; catalog # NB300–143, Novus Biologicals), and rabbit anti-amyloid-β (1:300; catalog # 71–5800, Invitrogen). .. Secondary antibodies included: goat anti-mouse AlexaFluor 488 secondary (1:500; catalog # A-11029, Invitrogen) and goat anti-rabbit AlexaFluor 594 secondary (1:500; catalog # A-11037, Invitrogen).
Immunohistochemistry:Article Title: Critical Role of GFR?1 in the Development and Function of the Main Olfactory System
Article Snippet: The signal was detected with an alkaline phosphatase-conjugated anti-DIG antibody (Roche) at 4°C followed by NBT/BCIP (Promega) solutions at room temperature. .. Immunohistochemistry Slides were removed from the − 20°C, air dried, blocked for 1 h (0.1 M PBS, 5% normal donkey serum, and 0.2% Triton X-100), and incubated in the following primary antibodies (same as block solution) overnight at room temperature: goat anti-GFRα1 (1;200;R&D), rabbit anti-GAP-43 (1;500; Novus Biological), goat anti-OMP (1;5000; Wako Pure Chemicals ), rabbit anti-cleaved caspase-3 (1;500; Cell Signaling Technology), mouse anti-Reelin clone G10 (1;300; Millipore), chicken anti-T-box brain (Tbr1) (1:250; Millipore), rabbit anti-calbindin (CB) (1:1000; Millipore), rabbit anti-calretinin (CR) (1:2000; Millipore), rabbit antityrosine hydroxylase (TH) (1:1000; Millipore), rabbit anti-laminin (LN) (1:500; Sigma), rat anti-integrin- α6 (1:500; BD Biosciences), rat anti-NCAM (1:500; BD PharMingen), and guinea pig anti-vesicular glutamate transporter 2 (VGlut2) (1:400; Millipore). .. After 3 (10 min) washes in PBS, slides were incubated with the following fluorescent secondary antibodies (same diluent as block) for 2 h at room temperature: donkey anti-goat Alexa Fluor 488, 568, or 647; donkey anti-rabbit Alexa Fluor 488, 555, or 647; donkey anti-mouse Alexa Fluor 488, 555, or 647 (all Alexa Fluor secondary’s from Invitrogen used at 1:500); donkey anti-rat DyLight 549; donkey anti-chicken DyLight 488; and donkey anti-guinea pig DyLight 549 (all DyLight secondary’s from Jackson ImmunoResearch used at 1:400).
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