Gb11373

Manufactured by Wuhan Servicebio Technology

Sourced in China

GB11373 is a laboratory equipment product. It is designed to perform a specific technical function, but without further details a concise and unbiased description cannot be provided while avoiding interpretation or extrapolation. Description not available.

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Lab products found in correlation

5 protocols using gb11373

Evaluating ECM, Inflammatory Markers, and Macrophages

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To evaluate in vivo changes in ECM secretion, levels of
inflammatory cytokines, and macrophage polarization, the sections were
immunohistochemically stained. Briefly, deparaffinized sections were
incubated with 3% H₂O₂ for 15 min in the dark and then
blocked with 3% BSA for 30 min at room temperature. The sections were
incubated overnight at 4°C along with the following primary antibodies: Col
II (1:500, GB111629, Servicebio), aggrecan (1:500, GB11373, Servicebio),
IL-1β (1:200, AF5103, Affinity), IL-6 (1:200, DF6087, Affinity), IL-10
(1:200, DF6894, Affinity), IL-4 (1:200, AF5142, Affinity), CD68 (1:300,
GB113109, Servicebio), Arg1 (1:500, GB11285, Servicebio), and CC-chemokine
receptor 7 (CCR7) (1:300, GB11502, Servicebio). After primary antibody
incubation, the sections were incubated with a specific HRP-conjugated
secondary antibody (1:200, S0001, Affinity) for 1 h at 25°C. The sections
were stained with 3,3′-diaminobenzidine and counterstained with hematoxylin
for the nucleus. Finally, the sections were mounted and photographed using
an optical microscope (Nikon, Tokyo, Japan). Immunohistochemical staining
intensity was digitally quantified using ImageJ software.

Li W., Zhou P., Yan B., Qi M., Chen Y., Shang L., Guan J., Zhang L, & Mao Y. (2023). Disc regeneration by injectable fucoidan-methacrylated dextran hydrogels through mechanical transduction and macrophage immunomodulation. Journal of Tissue Engineering, 14, 20417314231180050.

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Immunofluorescence Staining of Disc Cells

Cited 1 time

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Our previous study also described this method in detailly [2 (link)]. After intervertebral disc sections or NP cells were well prepared, 0.1% Triton X-100 was employed to permeate samples for 5 min. Then, samples were blocked with 3-5% BSA for 60 min at 37°C. Then, the samples were incubated using primary antibody against γH2AX (GB111841, 1 : 200, Servicebio, Wuhan, China), aggrecan (GB11373, 1 : 500, Servicebio, Wuhan, China), collagen type II (GB14073, 1 : 500, Servicebio, Wuhan, China), iNOS (GB11119, 1 : 1000, Servicebio, Wuhan, China), collagen type I (GB11022, 1 : 500, Servicebio, Wuhan, China), NLRP3 (GB11300, 1 : 1000, Servicebio, Wuhan, China), CD206 (#24595, 1 : 800, Cell Signaling Technology, Inc. USA), MMP3 (GB11131, 1 : 500, Servicebio, Wuhan, China), AGT (ab108334, 1 : 200, Abcam, USA), Nrf2 (340675, 1 : 500, Zenbio, Chengdu, China), and p65 (GB11142, 1 : 200, Servicebio, Wuhan, China) at 4°C overnight. At the second day, samples were treated with fluorescence secondary antibody (GB22303, GB21301, Servicebio, Wuhan, China) for 1 h in the dark room. The nuclei were staining with DAPI solution (G1012-100ML, Servicebio, Wuhan, China). The fluorescence was detected using fluorescence microscope (Olympus, Japan).

Sun K., Sun X., Sun J., Jiang Y., Lin F., Kong F., Li F., Zhu J., Huan L., Zheng B., Wang Y., Zou W., Gao L., Xu X, & Shi J. (2021). Tissue Renin-Angiotensin System (tRAS) Induce Intervertebral Disc Degeneration by Activating Oxidative Stress and Inflammatory Reaction. Oxidative Medicine and Cellular Longevity, 2021, 3225439.

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Immunohistochemical Analysis of Articular Tissue

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Paraffin sections of the articular tissues were subjected to IHC analysis. Briefly, after antigen retrieval, the samples were incubated with primary antibodies, including rabbit anti-MMP13 (1:250, GB11247, Servicebio, China), Col 2 (1:250, GB11021, Servicebio, China), and ACAN (1:500, GB11373, Servicebio, China) overnight at 4 °C, followed by binding with biotinylated secondary antibodies and incubation with diaminobenzidine (DAB) substrate for 10 min. The samples were imaged under the fluorescence microscope (ECLIPSE Ci-L, Nikon, Japan). The positive areas of MMP13, Col 2, and ACAN were quantified through ImageJ software.

Zhou D., Wei Y., Sheng S., Wang M., Lv J., Zhao B., Chen X., Xu K., Bai L., Wu Y., Song P., Cao L., Zhou F., Zhang H., Shi Z, & Su J. (2024). MMP13-targeted siRNA-loaded micelles for diagnosis and treatment of posttraumatic osteoarthritis. Bioactive Materials, 37, 378-392.

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Immunofluorescent Staining of NP Cells

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For aggrecan, type II collagen, MMP3, Ki-67, p65, and Nrf2 immunofluorescent staining, NP cells with different treatments were fixed using 4% PFA for 15-20 min and were permeated for another 5 min using 0.1% v/v Triton X-100. Cells were incubated with Aggrecan (GB11373, 1 : 500-1 : 1000, Servicebio, Wuhan, China), type II collagen (GB11021, 1 : 100-1 : 500, Servicebio, Wuhan, China), MMP3 (GB11131, 1 : 400-1 : 1600, Servicebio, Wuhan, China), Ki-67 (GB111141, 1 : 1200, Servicebio, Wuhan, China), NF-κB p65 (D14E12, #8242, cell signaling technology, Inc., 3 Trask Lane Danvers, USA), and Nrf2 (340675, Zenbio, Chengdu, China, 1 : 100) diluted in 0.2% w/v bovine serum albumin- (BSA-) TBS for 1 h and then washed with PBS. Cells were incubated with DAPI solution (G1012-100ML, Servicebio, Wuhan, China) for visualization of nuclei and, then, incubated with FITC-conjugated goat anti-rabbit IgG (GB22303, Servicebio, Wuhan, China) and Cy3-conjugated goat anti-rabbit IgG (GB21301, Servicebio, Wuhan, China) for 30 mins in the dark environment. Fluorescence detection was performed by fluorescence microscope (Olympus, Japan).

Luo X., Huan L., Lin F., Kong F., Sun X., Li F., Zhu J., Sun J., Xu X., Sun K., Duan L, & Shi J. (2021). Ulinastatin Ameliorates IL-1β-Induced Cell Dysfunction in Human Nucleus Pulposus Cells via Nrf2/NF-κB Pathway. Oxidative Medicine and Cellular Longevity, 2021, 5558687.

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Immunofluorescence Analysis of Chondrocytes

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Chondrocytes were fixed in 4% paraformaldehyde for 15 min, and washed with PBS three times. The normal goat serum (Servicebio, WGAR1009) was used to block chondrocytes for 30 min at room temperature. The chondrocytes were incubated with primary antibodies against aggrecan (1:500, Servicebio, GB11373) and collagen-II (1:200, Servicebio, GB11021) at 4 °C for 24 h. Then chondrocytes were incubated with fluorescein-conjugated goat anti-rabbit IgG (1:600, Servicebio, GB21301) at 37 °C for 1 h. The nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI). The images were acquired by an inverted fluorescence microscope. Cellular fluorescence intensity was quantified by evaluating positive cells using Image-J software.

Zhuang H., Ren X., Jiang F, & Zhou P. (2023). Indole-3-propionic acid alleviates chondrocytes inflammation and osteoarthritis via the AhR/NF-κB axis. Molecular Medicine, 29, 17.

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