Tissue sections were permeabilized with PBS/0.3% Tween-20 (PBST) for ~ 30 min then blocked with 5% normal donkey or normal horse serum in 0.01% PBST at room temperature for ~ 1 h. Primary antibodies were diluted in 0.01% PBST and 5% serum then incubated overnight at 4 °C with gentle shaking. Primary antibodies used for this study were; rabbit anti-TARDBP (Proteintech 10,782–2-AP), mouse anti-NEUN (Millipore Sigma MAB377), and DAPI (4′,6-diamidino-2-phenylindole)(Sigma Aldrich 10236276001). After rinsing in PBST 3X, secondary antibodies diluted in 0.01% PBST and 5% serum were added and incubated at room temperature for ~ 1 h with gentle shaking. Secondary antibodies included Alexa Fluor 555 anti-rabbit (Abcam ab150106) and Alexa Fluor 647 anti-mouse (Jackson ImmunoResearch 715-605-151). DAPI was used for nucleus staining. Imaging was performed using a Zeiss LSM 800 laser scanning confocal microscope (LSCM) with Airyscan.
Ab150106
Manufactured by Abcam
Sourced in United Kingdom
Ab150106 is a recombinant polyclonal antibody targeted against the human PD-1 protein. It is suitable for use in various immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Alexa fluor 647 anti mouse, by Jackson ImmunoResearch (1 mentions)
Mab377, by Merck Group (1 mentions)
Lsm 800, by Zeiss (1 mentions)
Ab53554, by Abcam (1 mentions)
Ab126649, by Abcam (1 mentions)
Ab2072, by Abcam (1 mentions)
Af555, by Thermo Fisher Scientific (1 mentions)
Axio scan z1 slide scanner microscope, by Zeiss (1 mentions)
Moab 2, by Abcam (1 mentions)
Mkb 2213, by Vector Laboratories (1 mentions)
Ab20066, by Abcam (1 mentions)
Ab181023, by Abcam (1 mentions)
Ab150075, by Abcam (1 mentions)
Sc 5303, by Santa Cruz Biotechnology (1 mentions)
Sp8 laser scanning confocal microscope, by Leica (1 mentions)
Ab150077, by Abcam (1 mentions)
3 protocols using ab150106
1
Immunofluorescence Staining of Neural Proteins
2
Multimodal Immunolabeling of Mouse Brain
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Mouse brain sections were blocked with MOM solution (#MKB-2213, Vector Labs,) and then incubated overnight with primary antibodies against INFγ (1:100, #MCA1301, BioRad), GFAP (1:1000, #ab53554, Abcam) and APP (1:200, #ab2072, Abcam) or against Aβ (MOAB-2 1:1000, #ab126649, Abcam). The next day, the sections were incubated with donkey anti-mouse (AF555, 1:500, #A3157, Thermo Fisher), donkey anti-goat (AF488, 1:500, #A11055, Thermo Fisher) and donkey anti-rabbit (AF647, 1:500, #A31573, Thermo Fisher) or donkey anti-mouse (AF555, 1:500, #ab150106, Abcam) secondary antibodies followed by DAPI staining and Mowiol mounting.
Sections stained with secondary antibodies only were used as negative controls. Additionally, bleed through control stainings were incubated with all three primary antibodies and one secondary separately. Stained mouse brain sections were imaged using a 10x objective on AxioScan.Z1 slide scanner microscope (Zeiss, Oberkochen). To better visualize the distribution of specific markers the slides were imaged also with a 63x oil-immersion objective on a LSM780 confocal microscope (Zeiss, Oberkochen).
Sections stained with secondary antibodies only were used as negative controls. Additionally, bleed through control stainings were incubated with all three primary antibodies and one secondary separately. Stained mouse brain sections were imaged using a 10x objective on AxioScan.Z1 slide scanner microscope (Zeiss, Oberkochen). To better visualize the distribution of specific markers the slides were imaged also with a 63x oil-immersion objective on a LSM780 confocal microscope (Zeiss, Oberkochen).
3
Immunohistochemical Characterization of Cholinergic and Dopaminergic Receptors
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Coronal slices were prepared and blocked for 1 h at room temperature in 1% albumin and 0.5% Triton-X in PBS. Slices were then incubated overnight at 4 °C in 1% albumin in PBS with the following antibodies: anti-ChAT (1:200, ab181023, Abcam, Cambridge, UK); antidopamine receptor 1 (D1R, 1:100, Ab20066, Abcam, Cambridge, UK) and anti-dopamine receptor 2 (D2R, 1:100, sc-5303, Santa Cruz, Dallas, TX, USA). Sections were then dark incubated 1 h at room temperature in blocking buffer with AlexaFluor 488 donkey anti-goat IgG (1:400, ab150077, Abcam, Cambridge, UK), AlexaFluor 555 donkey anti-mouse IgG (1:400, ab150106, Abcam, Cambridge, UK) and AlexaFluor 647 donkey anti-rabbit IgG (1:400, ab150075, Abcam, Cambridge, UK). Sections were washed with PBS (×3, 5 min each) after each incubation step. Samples were visualized using the Leica SP8 laser scanning confocal microscope and Leica LAS X image acquisi- tion software. Quantitative analysis (cell counts) was performed using LAS X software (Version 2.0.1, Mannheim, Germany).
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