Rabbit anti-human STAT3 monoclonal antibody (1:100; cat. no. BSM-52235R; BIOSS); Diaminobenzidine (DAB) color reagent (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.); poly-l-lysine coated slides (cat. no. AR1065; Wuhan Boster Biological Technology, Ltd.); HRP-conjugated AffiniPure goat anti-rabbit IgG testing kit (cat. no. BA1054; Wuhan Boster Biological Technology, Ltd.); Periodic Acid-Schiff Stain kit, (cat. no. DG0005; Beijing Leigen Biotechnology Co., Ltd.); 5% BSA (cat. no. AR0004; Wuhan Boster Biological Technology, Ltd.) and normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.)
Normal goat serum (ngs)
Manufactured by Wuhan Boster Biological Technology
Sourced in China
Normal goat serum is a biological reagent derived from the blood of healthy goats. It is used as a source of proteins, vitamins, and other biomolecules for various applications in research and laboratory testing.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Glial fibrillary acidic protein (gfap), by Abcam (1 mentions)
Image pro plus software 6, by Media Cybernetics (1 mentions)
Tcs sp2 confocal laser scanning microscope, by Leica (1 mentions)
Nmdar1, by Abcam (1 mentions)
In situ cell death detection kit, by Roche (1 mentions)
Confocal laser scanning microscope, by Olympus (1 mentions)
Sc 398675, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Axio imager a1, by Zeiss (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Wuhan Boster Biological Technology (1 mentions)
Ab175473, by Abcam (1 mentions)
Ab150141, by Abcam (1 mentions)
Sc 18796, by Santa Cruz Biotechnology (1 mentions)
Sc 293136, by Santa Cruz Biotechnology (1 mentions)
Te2000s inverted microscope, by Nikon (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Purmorphamine, by Selleck Chemicals (1 mentions)
Vismodegib, by Selleck Chemicals (1 mentions)
11 protocols using normal goat serum (ngs)
1
Immunohistochemical Analysis of STAT3
2
Immunohistochemical Analysis of Cerebral Cortex
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Four-micron frozen cerebral cortex sections fixed in acetone or 4% formaldehyde were blocked for endogenous peroxidase activity with 0.03% H2O2 if appropriate. Blocking was achieved with phosphate-buffered saline (PBS) containing 5% normal goat serum (Wuhan Boster Biological Technology, Ltd., Wuhan, China) for 1 h at room temperature. Sections were then incubated overnight at 4°C with the following primary antibodies; NMDAR1 (1:100; mouse monoclonal; Abcam, Cambridge, MA, USA), CaM (1:100; mouse monoclonal; Abcam), nNOS (1:50; Rabbit monoclonal; Abcam), sGC (1:50; rabbit polyclonal; Abcam), cGMP (1:50; mouse monoclonal; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and GFAP (1:50; rabbit polyclonal/mouse monoclonal; Abcam). Binding of primary antibodies was detected by incubating the sections for 30 min with fluorescein isothiocyanate (FITC) (green)/Alexa Fluor 594 (red) conjugated secondary antibody. Imaging was performed with a Leica TCS SP2 confocal laser scanning microscope (Leica Microsystems, Wetzlar, Germany). The image data were analyzed and quantified using ImagePro Plus software 6.0 (Media Cybernetics, Inc., Rockville, MD, USA).
3
Quantifying Apoptotic Endothelial Cells in Aorta
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For the detection and quantification of apoptotic endothelial cells in aortic sections, TUNEL technology (In Situ Cell Death Detection kit; Roche Applied Science, Madison, WI, USA) was used in combination with immunofluorescence for CD31. The paraffin-embedded sections were stained with a primary rabbit polyclonal anti-CD31 antibody (1:50 dilution; catalog no. ab28364; Abcam, Cambridge, UK) followed by incubation with a Texas Red-conjugated goat anti-rabbit IgG secondary antibody (1:100 dilution; catalog no. BA1032; Wuhan Boster Biological Technology, Ltd., Wuhan, China) to visualize the endothelial cell layer. Briefly, sections were blocked with normal goat serum (Wuhan Boster Biological Technology, Ltd.) for 30 min at room temperature prior to immunofluorescence staining. The sections were incubated with anti-CD31 antibody overnight at 4°C. Subsequently, after washing with PBS containing 0.05% Tween-20, the secondary antibody was applied to the sections for 1 h at 20–37°C. The aortic segments embedded in paraffin were then used to detect TUNEL-positive apoptotic cells, according to the manufacturer's protocol. The nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI). The percentage of apoptotic endothelial cells was calculated as the proportion of TUNEL/DAPI/CD31-positive cells out of total DAPI/CD31-positive cells.
4
Immunofluorescence Staining of MRTF-A
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Following transfection, the cells were fixed in 4% paraformaldehyde for 15 min at room temperature and then blocked with normal goat serum (Wuhan Boster Biological Technology, Ltd., Wuhan, China) for 20 min at room temperature. Following incubation with the primary antibody (cat. no. sc-398675, mouse anti-MRTF-A; 1:200; Santa Cruz Biotechnology, Inc.) in a humidified chamber overnight at 4°C, cells were incubated with the secondary antibody [cat. no. BA1101; fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG; 1:100; Wuhan Boster Biological Technology, Ltd.] for 30 min at 37°C. Subsequently, cells were incubated with DAPI (5 µg/ml; cat. no. C1005; Beyotime Institute of Biotechnology) for 15 min at room temperature. Following washing with PBS, the samples were observed under laser scanning confocal microscope (magnification, ×200; Olympus Corporation, Tokyo, Japan).
5
Quantifying PKM2 Expression in BGC-823 Cells
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BGC-823 cells were seeded at a density of 1×106 cells/well into 6-well plates, prior to treatment with LY294002 (0, 50 or 100 µmol/l) or with DMSO (0.2%) (2 µl/well) for 48 h at 37°C. Subsequent to washing with PBS 3 times, the cells were fixed with cold acetone for 10 min at 4°C. Next, the cells were blocked with 10% normal goat serum (Wuhan Boster Biological Technology, Ltd., Wuhan, China) for 30 min and probed with an antibody against PKM2 (cat. no. 4053, dilution, 1:100, Cell Signaling Technology, Inc.) at 4°C overnight. Alexa Fluor dye conjugated secondary antibodies (2 mg/ml goat anti-rabbit IgG (H+L) highly cross-absorbed) (cat. no. R37117, Alexa Fluor 594; dilution, 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.) were incubated with cells for 1 h at 20°C to enable the samples to be visualized under a fluorescent microscope (Axio Imager A1; Carl Zeiss AG, Oberkochen, Germany). The nuclei were stained using DAPI (2 µg/ml; Invitrogen; Thermo Fisher Scientific, Inc.) for 15 min under dark conditions at 20°C.
6
Histological Analysis of DRG Tissue in Chronic Sciatic Nerve Compression
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At 3 weeks following the establishment of the chronic sciatic nerve compression model, the contra-lateral and ipsilateral L4–L6 DRG tissues were removed, embedded in paraffin and sectioned. The samples were blocked with normal goat serum (Wuhan Boster Biological Technology, Ltd.) and probed with anti-TGF-β1 (1:50 dilution), anti-CTGF (1:50 dilution) or anti-collagen I (1:200 dilution) at 4°C overnight. The following day, the slides were washed three times with phosphate-buffered saline and incubated with secondary antibody at 4°C overnight, then washed with Tris-buffered saline plus Tween-20 (TBS-T; 0.1% Tween-20). Following washing, the nuclei were counterstained with DAPI (100 µg/ml; Wuhan Boster Biological Technology, Ltd.), and samples were mounted in antifade reagents (Beyotime Institute of Biotechnology, Haimen, China). The fluorescence was examined under a fluorescent microscope (Olympus IX71; Olympus Corporation).
7
Immunohistochemical Analysis of CD163 and Co-Expressed Markers in Liver Tissues
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Liver tissues were fixed in formalin and blocked in 10% normal goat serum (Wuhan Boster Biological Technology, Ltd., Wuhan, China) for 30 min at 37°C. The tissues were incubated with a primary antibody against CD163 (1:100; sc-18796; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) overnight at 4°C and amplified with rabbit anti-goat IgG secondary antibody (1:1,000; ab150141; Abcam, Cambridge, England) conjugated to Alexa Fluor 488 for 1 h at 37°C. For co-localization with CD163, sections were co-stained overnight with a primary antibody against p-JNK (1:100; sc-293136) or MIP-1α (1:100; sc-365691) (both from Santa Cruz Biotechnology, Inc.) overnight at 4°C and amplified with goat anti-mouse IgG secondary antibody (1:500; ab175473; Abcam) conjugated to Alexa Fluor 568 for 1 h at 37°C. Prior to observation, DAPI was used to label the nuclei. Images were acquired using a fluorescence microscope (OLYMPUS BX51; Olympus Corporation, Tokyo, Japan) with appropriate filters. The images are presented as single-color stains of green and red to exhibit the localization of the two markers in the cells. Merged images are presented below the images with single-color stains. The two markers that were co-expressed in cells at a similar location are often observed in yellow.
8
Immunofluorescence Assay for p53 and p21
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For determination of the distribution of p53 and p21, cells were fixed in ice-cold 4% (w/v) paraformaldehyde in phosphate-buffered saline (PBS) for 30 min and permeabilized with 0.2% Triton Χ-100 (Shanghai Chemical Reagent Co., Ltd.) for 10 min at 40°C. Cells were treated with 3% H2O2 for 20 min and non-specific binding was blocked with 10% normal goat serum (Wuhan Boster Biological Technology, Ltd., Wuhan, China) for 30 min at 37°C. Following washing with PBS three times, cells were incubated with mouse monoclonal anti-p53 (1:200; cat. no. AP062; Beyotime Institute of Biotechnology, Haimen, China) and rabbit polyclonal p21 antibodies (1:200; cat. no. sc-397; Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4°C followed by 1 h incubation with goat anti-rabbit (1:64; cat. no. BA1105; Boster Biological Technology, Ltd.) and goat anti-mouse fluorescein isothiocyanate-conjugated (1:64; cat. no. BA1101; Boster Biological Technology, Ltd.) antibodies. Following counterstaining with DAPI (Invitrogen Life Technologies, Carlsbad, CA, USA), cells were visualized under the Nikon TE2000-S inverted microscope (Nikon, Tokyo, Japan).
9
Hedgehog Pathway Inhibitors in Cell Studies
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Dimethyl sulfoxide (DMSO) was obtained from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Vismodegib (cat no. s1082) and Purmorphamine (cat no. s3042) was obtained from Selleck Chemicals (Houston, TX, USA). Lipofectamine® 3000 was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Antibody against Gli1 (cat no. sc-20687) was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Antibody against EDNRB (cat no. ab117529) was purchased from Abcam (Cambridge, MA, USA). Antibody against GAPDH (cat no. 10494-1-AP) was purchased from Wuhan Sanying Biotechnology (Wuhan, China). The secondary goat anti-rabbit antibody (cat no. 7074S) was purchased from Cell Signaling Technology (Danvers, MA, USA). The H&E staining kit (cat no. AR1180), immunohistochemical SABC reagent kit (cat no. SA1020), DAB (cat no. AR1022) and normal goat serum (cat no. AR0009) were purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).
10
Immunohistochemical Analysis of Intestinal Tight Junction Proteins
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The ileal tissue specimens were cut into 10 µm sections following dewaxing and hydrating. Sections were treated with 3% H2O2-methanol to block endogenous peroxidase activity, following which they were incubated with 5% normal goat serum (Wuhan Boster Biological Technology, Ltd.) at room temperature for 10 min and incubated with ZO-1 and occludin antibodies (dilution 1:200) overnight at 4°C. Slides were subsequently washed with PBS and incubated with a biotinylated secondary antibody (1:500; cat. no. SA00004-2; Wuhan Boster Biological Technology, Ltd.) for 1 h at room temperature. Slides were washed with PBS again and incubated with HRP-labeled streptavidin (1:200; ab214880; Abcam) at 37°C for 1 h. Samples were developed using diaminobenzene (DAB) at room temperature for 30 sec and counterstained with hematoxylin at room temperature for 5 min. Slides were rinsed in distilled water and dehydrated, following which they were observed under a light microscope (magnification, ×100).
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