Frozen sectioning was used to slice brain samples from AD patients and non-AD patients; samples were donated by the Department of Anatomy, Histology, and Embryology (School of Basic Medicine, Fudan University, Shanghai, China). Sections (20 μm) were rinsed three times with PBS and then permeabilized with 0.1% Triton X-100 in PBS, blocked with 5% BSA in PBS at RT for 30 min, and incubated overnight at RT with both rabbit SV2A antibody (1 μg/ml, Abcam) and mouse APP antibody (1 μg/ml, Covance). Subsequently, sections were washed three times in PBS, followed by a 2-h incubation with goat anti-rabbit IgG (594; 3 μg/ml, Abcam) and goat anti-mouse IgG (488; 3 μg/ml, Abcam). Sections were again washed three times with PBS. Fluorescence intensity was detected by using a Zeiss LSM710 fluorescence microscope (Liu et al., 2018 (link)).
Goat anti rabbit igg 594
Manufactured by Abcam
Sourced in United States
Goat anti-rabbit IgG 594 is a secondary antibody conjugated with the fluorescent dye Alexa Fluor 594. It is designed to recognize and bind to rabbit IgG antibodies, allowing for the detection and visualization of target proteins in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit sv2a antibody, by Abcam (1 mentions)
Lsm710 fluorescence microscope, by Zeiss (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Ab96901, by Abcam (1 mentions)
M0851, by Agilent Technologies (1 mentions)
Goat anti mouse igg 488, by Thermo Fisher Scientific (1 mentions)
A11001, by Thermo Fisher Scientific (1 mentions)
Tcs sp8, by Leica (1 mentions)
Las af, by Leica (1 mentions)
Monoclonal mouse anti vimentin, by Agilent Technologies (1 mentions)
3 protocols using goat anti rabbit igg 594
1
Quantifying Alzheimer's Pathology in Brain Samples
2
Quantification of Phosphorylated p38 and JNK in Penile Smooth Muscle Cells
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To assess the number of SM cells positive for phosphorylated p38 or phosphorylated JNK, paraffin-embedded sections (2.5-mm-thick) of penile tissue were incubated overnight with primary antibodies against α-SMA (a SM cell marker, 1:100, M0851, Dako, Glostrup, Denmark; red) and phosphorylated p38 (1:20, #4511, Cell-Signaling Technology, Danvers, MA, USA; green) or phosphorylated JNK (1:20, #4668S, Cell-Signaling Technology, Danvers, MA, USA; green). After washing in PBS, the sections were incubated with two secondary antibodies (goat anti-mouse IgG 488 [1:400, A-11001, Invitrogen, Camarillo, CA, USA] and goat anti-rabbit IgG 594 [1:200, ab96901, Abcam, Cambridge, CB4 0FL, UK]) in 1% bovine serum albumin at room temperature for 1 h. Digital images were acquired using a confocal microscope (Leica TCS SP8, Leica Microsystems, Wetzlar, Germany). Ten rats from each group (two sections per animal) were analyzed. Under confocal microscopy, the number of SM cells positive for phosphorylated p38 or phosphorylated JNK (yellow) was quantified from five randomly selected high-power fields (white arrow). The slides were evaluated by three independent observers in a blinded fashion.
3
Co-localization of Fibroblasts and Signaling Proteins
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To assess co-localization of fibroblasts and ROCK1 or LIMK2-activated sites in penile tissue, double immunofluorescence for ROCK1 and vimentin or phospho-LIMK2 and vimentin were performed. For immunofluorescence staining of paraffin embedded sections (2.5µm) were treated and incubated with primary antibody; monoclonal mouse anti-vimentin (1:100, Dako, Glostrup, Denmark) during two overnight periods, monoclonal rabbit anti-ROCK1 (1:10, AbCam, Cambridge, MA, USA) or rabbit polyclonal anti-LIMK2 (phospho T505) (1:100, AbCam, Cambridge, MA, USA) for two overnight periods at room temperature. After three washes in PBS, the sections were incubated with the fluorescence secondary antibody; goat antimouse IgG antibody 488 (1:400, Invitrogen, Camarillo, CA, USA) 10min for vimentin, and goat anti-rabbit IgG 594 (1:200, AbCam, Cambridge, MA, USA) 10min for ROCK1 or phospho-LIMK2. Sections were analyzed using a confocal laser fluorescencemicroscope (Leica Microsystems, Heidleberg, Germany) with single and double filter settings at 488 and 594 nm.
Images were acquired using LAS AF (Leica Microsystems, Heidleberg, Germany).
Images were acquired using LAS AF (Leica Microsystems, Heidleberg, Germany).
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