MEC-1 and JVM-3 were plated on slides in 48-well plates and incubated at 60°C for IF and RNA-FISH analysis. After fixation and permeabilization, cells were incubated overnight at 4°C with primary diluted antibody anti-Ki67 (#ab15580, Abcam). The secondary antibody Alexa Fluor® 594 - Conjugated Goat anti-Rabbit IgG (H+L) was purchased from ZSGB-BIO, China. Fluorescence were observed using Axio Scope.A1 (Zeiss). FISH assay was performed using Fluorescent In Situ Hybridization Kit (RiboBio, Guangzhou, China) according to the manufacturer’s guidelines. And Cy3-labeled probes from RiboBio, Guangzhou, China were measured by visualized with a confocal microscopy.
Alexa fluor 594 conjugated goat anti rabbit igg h l
Manufactured by ZSGB-BIO
Sourced in China
Alexa Fluor® 594 - Conjugated Goat anti-Rabbit IgG (H+L) is a secondary antibody conjugated with the fluorescent dye Alexa Fluor® 594. It is designed to detect and visualize rabbit primary antibodies in various immunoassays and imaging applications.
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3 protocols using alexa fluor 594 conjugated goat anti rabbit igg h l
1
Immunofluorescence and RNA-FISH Analysis
2
Immunofluorescence Staining of YAP1
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Cells cultured on glass coverslips were fixed with 4% paraformaldehyde followed by permeabilization with 0.1% Triton X‐100. Subsequently, cells were blocked with 1% BSA and incubated with YAP1 antibody, washed with PBS, incubated with Alexa Fluor 594‐Conjugated Goat anti‐Rabbit IgG (H + L) (#ZF‐0516, ZSGB‐BIO) as the secondary antibody, and counterstained with DAPI (#C1002, Beyotime Biotechnology). Images were taken at 60X magnification.
3
Renal Tissue Immunofluorescence Analysis
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Immuno uorescence stainings of Fractalkine, p-p38, and Foxp3 proteins were performed on 4 µm-thick para nembedded sections of renal tissues after incubation with a primary antibody targeting Fractalkine (DF12376, 1:80; A nity), Foxp3 (GTX107737, 1:100; GENE TEX), or p-p38 (AF4001, 1:80; A nity) overnight at 4 °C. Next, the samples were incubated with Alexa Fluor 594-conjugated goat anti-rabbit IgG (H + L) (ZF0516, 1:100; ZSGB-BIO) for 1 h at 24 °C. Then, the samples were observed with a uorescence microscope (IX71, Olympus, Tokyo, Japan) equipped with ISC capture software, and the images were collected with a CCD camera (Discovery C15, Olympus, Tokyo, Japan).
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