Paraffin-embedded sections were cut into 4um and deparaffinized in xylene and rehydrated, and endogenous peroxidase was blocked with 3%H2O2. Antigen retrieval was accomplished by 0.01 mol/L citrate buffer solution (pH 6.0) in a 700 W microwave oven for 15 min. After incubation with 5% normal goat serum for 20 min, the slides were exposed overnight at 4 °C to the rabbit anti-DEC2 (1:150; Proteintech), rabbit anti-Ki-67(1:800; Proteintech), rabbit anti-NR2F1 (1:200; Proteintech). Sections were then incubated with biotinylated goat anti-rabbit IgG (Zhongshan Goldenbridge Biotechnology) for 1 h, and streptavidin-peroxidase for 30 min. The 0.02% diaminobenzidine tetrahydrochloride was used as a chromogen, and the slides were counterstained with hematoxylin. The percentage of positive cells was estimated using an image analysis system (Leica).
Biotinylated goat anti rabbit igg
Manufactured by Zhongshan Golden Bridge Biotechnology
Sourced in China
Biotinylated goat anti-rabbit IgG is a laboratory reagent used to detect and bind to rabbit immunoglobulin G (IgG) molecules. It is a conjugate of biotin, a small molecule, and an antibody derived from goats that specifically recognizes and binds to rabbit IgG. This reagent is commonly used in various immunoassay techniques, such as Western blotting and enzyme-linked immunosorbent assays (ELISAs), to facilitate the detection and visualization of rabbit IgG-containing samples.
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Lab products found in correlation
4 protocols using biotinylated goat anti rabbit igg
1
Immunohistochemical Analysis of DEC2, Ki-67, and NR2F1
2
Immunohistochemical Analysis of Fas and FasL
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After 24 hours of reperfusion, rats were anesthetized with 10% (v/v) chloral hydrate and then fixed in 4% (w/v) paraformaldehyde through cardiac perfusion. The right hemisphere was harvested and sliced into paraffin-embedded sections at a thickness of 5 μm. Slices were blocked with normal goat serum (1:10, Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China), and incubated with rabbit anti-rat Fas and FasL polyclonal antibodies (1:100, Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.) at 4°C overnight; and then incubated with biotinylated goat anti-rabbit IgG (1:300, Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.) for 12 minutes at room temperature. Horseradish peroxidase-conjugated streptavidin working solution (1:300) was added and incubated for a further 12 minutes. Negative controls were incubated with PBS instead of antibodies. Three random sections from each rat were examined at five different visual fields under 200× magnification. The MiVnt image analysis system was used to count the number of Fas- and FasL-positive cells in the right hippocampus. The results were averaged.
3
Quantifying p-p38MAPK-Positive Cells in Rat Brain Slices
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Rat brain tissue slices were prepared, blocked with serum, and incubated with rabbit anti-rat p-p38MAPK monoclonal antibody (1:100; Cell Signaling Technology, Inc., Danvers, MA, USA) at 4°C for 24 hours, then with biotinylated goat anti-rabbit IgG (ready-to-use; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) at 37°C for 10-15 minutes, and finally with horseradish peroxidase-conjugated streptavidin. Subsequently, slices were developed with DAB, cleared with xylene, and mounted and observed under a light microscope (Olympus, Tokyo, Japan). Negative control slices were treated with phosphate-buffered saline rather than primary and secondary antibodies. The number of p-p38MAPK-positive cells was counted in each of two fields of vision for each slice, in five slices from each animal, under 200× magnification. The average value was obtained and grayscale analysis was performed using the Advance3.2 system (Motic Medical Laboratory, Xiamen, Fujian Province, China).
4
Immunohistochemical Analysis of HCC Xenograft Samples
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The study was approved by the Research Ethics Committee of Xi'an Jiaotong University. HCC samples from mice xenograft were examined immunohistochemically by treptavidin-peroxidase (SP) method, as described previously [56 (link)]. Briefly, paraformaldehyde-fixed paraffin embedded tissues were cut into 4-μm sections. The sections were dewaxed, dehydrated, following antigen retrieval in a pressure cooking for 5 min then incubated with 0.3% (v/v) hydrogen peroxide for 10 min to block non-specific antigens and any endogenous enzyme. The sections were incubated with normal goat serum for 10 min then with the primary antibody against NICD1, P21 or Bcl-2 at 4°C overnight. The slides were washed with PBS and incubated with biotinylated goat anti-rabbit IgG (1:100 dilution; Zhongshan Golden Bridge Biotechnology, Beijing, China) for 20 min as the secondary antibodies. Each slide was colored with DAB (Sigma, St. Louis, MO, USA) in a dark room before incubated with peroxidase-conjugated streptavidin (Zhongshan Golden Bridge Biotechnology) for 10 min. At last, all the sections were rinsed with running water, counterstained by hematoxylin and dehydrated in graded ethanol. The slides were read and marked by two independent pathology experts under a microscope (Olympus Optical Co, Tokyo, Japan). Haematoxylin and eosin (H&E) staining was performed according to regular procedures.
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