Cell cultures were prepared and fixed in 4% paraformaldehyde. Sciatic nerve segments were dissected, fixed in 4% paraformaldehyde, dehydrated in 30% sucrose at 4 °C, then cut and mounted onto microscope slides. Sciatic nerve sections and cell cultures were blocked with 5% goat serum for 30 min, incubated with primary antibodies overnight at 4 °C, and then incubated with fluorescent-dye-conjugated secondary antibodies for 1 h at room temperature. Primary antibodies included mouse anti-S100 antibody (1:1000 dilution, Sigma), rabbit anti-Ki67 antibody (1:200 dilution, Sigma), mouse anti-NF200 antibody (1:200 dilution, Sigma), rabbit anti-S100 antibody (1:200 dilution, Abcam), and Hoechst 33342 (1:5000 dilution, Life Technologies). Secondary antibodies included goat anti-mouse IgG-Alex-488 (1:500 dilution, Abcam), and goat anti-rabbit IgG-Cy3 (1:100 dilution, Abcam). Images were obtained under a fluorescence microscopy (Axio Imager M2, Carl Zeiss Microscopy GmbH, Jena, Germany).
Rabbit anti ki67 antibody
Manufactured by Merck Group
The Rabbit anti-Ki67 antibody is a primary antibody used in immunohistochemistry and immunocytochemistry applications to detect the presence of the Ki67 protein, which is a cellular marker for proliferation. It recognizes the Ki67 nuclear antigen and is useful for identifying and quantifying proliferating cells in various tissues and cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti nf200 antibody, by Merck Group (2 mentions)
Rabbit anti s100 antibody, by Abcam (2 mentions)
Axio imager m2, by Zeiss (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Goat anti mouse igg alex 488, by Abcam (1 mentions)
Mouse anti s100 antibody, by Merck Group (1 mentions)
Goat anti rabbit igg cy3, by Abcam (1 mentions)
Goat anti cd34 antibody, by R&D Systems (1 mentions)
Sheep anti rabbit igg cy3, by Abcam (1 mentions)
Donkey anti goat igg alex 488, by Abcam (1 mentions)
Fitc labeled gsl 1 isolectin b4, by Vector Laboratories (1 mentions)
Rat anti mouse cd206, by BioLegend (1 mentions)
Rabbit anti mouse f4 80, by Abcam (1 mentions)
F4 80, by BioLegend (1 mentions)
4 protocols using rabbit anti ki67 antibody
1
Immunofluorescence Analysis of Sciatic Nerve
2
Fluorescence Immunohistochemistry of Tissue Slices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tissue slices were blocked with 5% goat serum for 1 h at 37 °C, incubated with primary antibodies overnight at 4 °C, and then secondary antibodies for 1 h at room temperature. Primary antibodies included goat anti-CD34 antibody (1:50 dilution, R&D), rabbit anti-S100 antibody (1:200 dilution, Abcam), rabbit anti-Ki67 antibody (1:200 dilution, Sigma), mouse anti-NF200 antibody (1:200 dilution, Sigma). Secondary antibodies included donkey anti-goat IgG-Alex-488 (1:500 dilution, Abcam), sheep anti-rabbit IgG-Cy3 (1:1000 dilution, Abcam) and donkey anti-mouse IgG-Alex-594 (1:800 dilution, Invitrogen). Nucleus was marked using Hoechst 33,342 (1:5000 dilution, Life Technologies). Images were acquired under a fluorescence microscopy (AxioImager M2, Zeiss).
3
Labeling Retinal and Choroidal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Retina or choroid membrane samples were fixed overnight at 4°C with 4% paraformaldehyde, blocked, and permeabilized in PBS containing 1% BSA and 0.3% Triton X‐100, then immunostained with FITC‐labeled GSL I‐isolectin B4 (Vector Laboratories) or rabbit anti‐Ki67 antibody (Millipore) at 4°C overnight. After washing, the samples were incubated with a cyanine 3–conjugated goat antirabbit IgG secondary antibody (Sigma‐Aldrich). Images were captured with a fluorescence or confocal microscope.
4
Immunofluorescence Validation of 4T1 Tumor
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ex vivo immunofluorescence staining of 4T1 tumor or normal tissues was performed to validate the results of in vivo studies. The experiments were performed, and the staining results were analyzed by two investigators (D. Gao and C. Zhang) who were blinded to the results of in vivo NIRF imaging and tumor therapy.
For CD206 and F4/80 overlay staining, 4T1 tumor, muscle, liver, or spleen tissue slices were incubated with rat anti-mouse CD206 (BioLegend, San Diego, CA) and rabbit anti-mouse F4/80 (Abcam, Cambridge, MA) primary antibodies for 1 h at room temperature, and then visualized with Cy3- or FITC-conjugated secondary antibodies under a confocal microscope. For Ki67 staining, the tumor slices were incubated with rabbit anti-Ki67 antibody (Millipore, Billerica, MA). The slices were then visualized with Cy3-conjugated secondary antibody under a confocal microscope. After staining, 10 random views in the tumor slices were selected for the analyses. The murine CD206 expression level was calculated by measuring the integrated optical density of images of an equivalent area using a previously described method 23 (link). The tumor cell proliferation index was calculated as the percentage Ki67-positive cells 24 (link).
For CD206 and F4/80 overlay staining, 4T1 tumor, muscle, liver, or spleen tissue slices were incubated with rat anti-mouse CD206 (BioLegend, San Diego, CA) and rabbit anti-mouse F4/80 (Abcam, Cambridge, MA) primary antibodies for 1 h at room temperature, and then visualized with Cy3- or FITC-conjugated secondary antibodies under a confocal microscope. For Ki67 staining, the tumor slices were incubated with rabbit anti-Ki67 antibody (Millipore, Billerica, MA). The slices were then visualized with Cy3-conjugated secondary antibody under a confocal microscope. After staining, 10 random views in the tumor slices were selected for the analyses. The murine CD206 expression level was calculated by measuring the integrated optical density of images of an equivalent area using a previously described method 23 (link). The tumor cell proliferation index was calculated as the percentage Ki67-positive cells 24 (link).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!