The experiment was performed in accordance with the manufacturer’s instructions; the SPlink Detection Kit was purchased from Zsgb-Bio Co. Ltd (Beijing, China). Renal tissues were fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned at a thickness of 5 µm. Antigen retrieval was carried out by boiling the samples in citrate buffer for 15 min at 92–98 °C; then, the sections were treated with 3% hydrogen peroxide for 30 min to inhibit peroxidase activity and were rinsed with PBS three times. Next, 10% normal goat serum was added for 1 h at 37 °C, and subsequently, incubation was carried out with anti-Mcp-1 (1:50, Abcam, Eugene, USA), anti-Tnf-α (1:50, BBI Life, Shanghai, China), and anti-Gal-3 (1:50, Santa Cruz Biotechnology, USA) overnight at 4 °C. The sections were then incubated with biotin-labeled secondary antibody at room temperature for 30 min. The nuclei were counterstained with hematoxylin. The immunohistological staining for Mcp-1, Tnf-α, and Gal-3 was observed by light microscopy.
Anti gal 3
Manufactured by Santa Cruz Biotechnology
Sourced in China, United States
Anti-Gal-3 is a laboratory reagent produced by Santa Cruz Biotechnology. It is an antibody that specifically binds to the Galectin-3 (Gal-3) protein. Gal-3 is a member of the lectin family and plays a role in various biological processes. The Anti-Gal-3 antibody can be used in research applications that involve the detection and study of Gal-3 expression or function.
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Lab products found in correlation
5 protocols using anti gal 3
1
Immunohistological Analysis of Renal Biomarkers
2
Localization of Gal-3 in Lung Tissue
3
Immunofluorescence Imaging of Cytokines
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Cells were tiled in a 24-well plate containing sterile cover glass slides and were cultured for 48 h, fixed with paraformaldehyde for 30 min, incubated with 0.3% Triton X-100 for 10 min, blocked with 3% goat serum for 1 h, and then incubated overnight with anti-Mcp-1 (1:50, Abcam, Eugene, USA), anti-Tnf-α (1:50, BBI Life, Shanghai, China), and anti-Gal-3 (1:50, Santa Cruz Biotechnology, USA). Then, cells were washed three times with PBS; DyLight488 or DyLight549 (Thermo Fisher Scientific, MA, USA) was then added into the cells for 1 h under dark conditions. Next, cells were washed with PBS three times and treated with DAPI for 10 min. Finally, the images were observed with a confocal microscope and were analyzed with LAS AF Lite (Leica, Solms, Germany).
4
Immunohistochemical Analysis of Gal-3 and Ki67 in Tumor Samples
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Paraffin-embedded tissue samples from all study patients were examined using hematoxylin and eosin staining to confirm tumor grade and type. Only one sample at the time of surgery was obtained for patients in adjuvant setting while two samples were taken from neoadjuvant patients; one at the time of diagnosis (Pre) and a second at the time of surgery after chemotherapy (Post). Gal-3 and Ki67 expression was detected by IHC using mouse polyclonal anti-Gal-3 (Santa Cruz, CA, USA, SC-14364, 1 : 500) and rabbit polyclonal anti-Ki67 (Abcam, Cambridge, MA, USA, ab92742, 1 : 2500) antibodies, respectively. After standard deparaffinization, samples were tested using Envision dual link dakocytomation kit following the manufacturer's protocol (Dako-Behring, Glostrup, Denmark). An irrelevant IgG was used as the primary antibody in controls. Scoring was performed by three blinded observers: two researchers and one pathologist in 3 different fields (at 200x and 400x magnification) for each specimen. Mean score by the three observers was recorded as the final result.
5
Immunohistochemical Analysis of Tumor Markers
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Paraffin-embedded tissue samples from all study patients were examined using hematoxylin and eosin staining to confirm tumor grade and type. Only one sample at the time of surgery was obtained for patients in adjuvant setting while two samples were taken from neo-adjuvant patients; one at the time of diagnosis (Pre) and a second at the time of surgery after chemotherapy (Post). Gal-3 and Ki67 expression was detected by IHC using mouse polyclonal anti-Gal-3 (Santa Cruz, CA, USA, SC-14364, 1:500) and rabbit polyclonal anti-Ki67 (abcam, Cambridge, MA, USA, ab92742, 1:2500) antibodies respectively. After standard deparaffinization, samples were tested using Envision dual link dakocytomation kit following the manufacturer's protocol (Dako-Behring, Glostrup, Denmark). An irrelevant IgG was used as the primary antibody in controls. Scoring was performed by three blinded observers; two researchers and one pathologist in 3 different fields (at 200x and 400x magnification) for each specimen. Mean score by the three observers was recorded as the final result.
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