Naloxone was purchased from Taizhou Hongyao Chemical Co., Ltd. Naltrexone and phloroglucinol were supplied by Aladdin. The dialysis bags, FITC, fura-2AM fluorescent probe, nissl staining solution, hematoxylin eosin (HE) staining kit and BCA protein concentration determination kit were ordred from Beyotime Biotechnology . LPS, Fetal bovine serum (FBS), DMEM, penicillin-streptomycin (PS) and trypsin-EDTA (0.25%) were purchased from Gibco Company. Protein lysate (RIPA), phenylmethanesulfonyl fluoride (PMSF) and 4′,6-diamidino-2-phenylindole (DAPI) were bought from Beijing Solarbio Science & Technology Co., Ltd. Anti-CD11b, anti-F4/80, anti-iNOS, anti-TNF-α, anti-IL-1β, anti-IL-6, anti-IL-10, anti-Bax, anti-caspase 3, anti-bcl-2, anti-β-tubulin, anti-GAPDH, anti-β-actin were supplied by Abcam.
Anti il 10
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-IL-10 is a laboratory reagent used for research purposes. It is an antibody that specifically binds to the cytokine interleukin-10 (IL-10). IL-10 is an important immunoregulatory molecule. The Anti-IL-10 antibody can be used to detect and study IL-10 in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Anti il 6, by Abcam (7 mentions)
Anti il 1β, by Abcam (6 mentions)
Anti tnf α, by Abcam (6 mentions)
Anti β actin, by Abcam (4 mentions)
Anti bax, by Abcam (3 mentions)
Anti inos, by Abcam (3 mentions)
Anti tgf β, by Abcam (3 mentions)
Anti bcl 2, by Abcam (2 mentions)
Hematoxylin eosin staining kit, by Beyotime (2 mentions)
Anti gapdh, by Abcam (2 mentions)
Anti β tubulin, by Abcam (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Malondialdehyde mda content assay kit, by Beyotime (2 mentions)
Total glutathione gsh px content assay kit, by Beyotime (2 mentions)
Superoxide dismutase sod content assay kit, by Beyotime (2 mentions)
Anti hif 1α, by Beyotime (2 mentions)
Anti foxp3, by Abcam (2 mentions)
Anti il 1β, by Santa Cruz Biotechnology (2 mentions)
Anti il 17, by Abcam (2 mentions)
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
30 protocols using anti il 10
1
Neuroinflammation and Apoptosis Assay
2
In Vivo Biocompatibility Assessment of Hydrogels
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All the animal experiments have been approved by the ethics committee of Shenzhen Top Biotech Co. Ltd. (no. TOP-IACUC-2020-09-0007). Hydrogel samples were implanted subcutaneously in rats to evaluate biocompatibility in vivo. After the rats were anesthetized with 1.5% isoflurane, the dorsal region was disinfected, and an incision of about 2 cm was made. A sterile hydrogel cube of 1 cm in length and width and 3 mm in thickness was implanted. The incision was sutured with 4-0 sutures. After implantation for 4 weeks, the rats were anesthetized and killed. The back hydrogel and surrounding tissues were taken out together and fixed with 4% paraformaldehyde. After dehydration, they were embedded in paraffin and cut into pathological sections. The sections were stained with hematoxylin and eosin (HE), and immunohistochemical staining was applied to quantify the inflammatory factors such as CD68, interleukin (IL)-10, and tumor necrosis factor-alpha (TNF-α). Immunohistochemical staining was performed as previously reported[39 (link)]. The primary antibodies used in this experiment were anti-collagen I, anti-collagen III, anti-IL-10, anti-TNF-α, and anti-CD68 antibodies (Abcam, Cambridge, MA, UK).
3
Western Blot Analysis of Protein Markers
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Western blot analysis was carried out as described previously.31 Protein concentrations were quantified using a Bio‐Rad assay (Bio‐Rad Laboratories, CA, USA). Proteins were resolved in a 10% SDS polyacrylamide gel and transferred to an Immobilon PVDF membrane (Amersham, Arlington Heights, IL, USA). Primary anti‐IL‐10 (Abcam, Cambridge, UK), anti‐E‐cadherin (BioLegend, San Diego, CA, USA), and anti‐GAPDH (Abcam) antibodies were used to sequentially probe the membrane.
4
CMV Infection and Inflammatory Response Analysis
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One day after CMV infection with or without Y27632 or K115, the cells were collected in RIPA buffer (Thermo Fisher Scientific) containing protease inhibitors (Roche Diagnostics, Basel, Switzerland). They were then sonicated and centrifuged. Western blotting (WB) was performed as described previously.34 (link) The primary antibodies were as follows: anti-TNF-α (1:500; Abcam), anti-NF-κB (1:100; Sigma-Aldrich), anti-IL-10 (1:500; Abcam), Phospho-IκBα (Ser32/36) (5A5) antibody (1:1000, Cell Signaling Technology), and anti-GAPDH (1:1000; Wako Pure Chemical Industries, Osaka, Japan). Horseradish peroxidase (HRP)-conjugated secondary antibody (1:2000) was purchased from Thermo Fisher Scientific. The bands were quantified using ImageJ 1.49 software (National Institutes of Health, Bethesda, MD, USA).
5
Cell Proliferation Assay with CCK8
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The cell proliferation ability was determined using the Cell Counting Kit-8 (CCK8, Dojindo, Japan). Briefly, 3000 cells were seeded into 96-well plates (Corning, USA). At the indicated times, each well medium was replaced with an equal volume of fresh medium containing 10% CCK-8 reagent. After cells were incubated at 37 °C for 2.5 h, absorbance was measured using an Enzyme-linked immunosorbent instrument (Thermo-Fisher, USA) at 450 nm. Recombinant human interleukin 10 (rhIL-10, R&D Systems, USA) and its neutralizing antibody (Anti-IL-10, Abcam, UK) were used in this study.
6
Immunofluorescence Staining of Lung Tissue
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Cell slides were de-paraffinized, rehydrated with an ethanol gradient, and 50% TritonX was added dropwise to the tissue and left for 10 min at room temperature. They were then washed with PBS thrice with 5 min each time. Citrate buffer was added to the slide and antigen retrieval was carried out at high temperature for 5 min. The slides were washed with 0.01 M PBS and closed with goat serum for l0 min at room temperature. Sections were incubated overnight at 4 °C with primary antibodies anti-CD86 (1:50; BIOSS, Beijing, China), anti-IL-6 (1:50; BIOSS, Beijing, China), anti-CD206 (1:50; Abcam, Shanghai, China), anti- IL-10 (1:50; Abcam, Shanghai, China). Unbound primary antibodies were washed off with PBS, and sections were incubated with secondary antibodies followed by nuclear staining and image acquisition. Lung tissue sections were incubated with primary antibodies anti-CD80 (1:50; BOSTER, Wuhan, Hubei, China), anti-IL-1β (1:100; BOSTER, Wuhan, Hubei, China), anti-IL-10 (1:50; BOSTER, Wuhan, Hubei, China) and anti-F4/80 (1:100; BOSTER, Wuhan, Hubei, China). Subsequent steps were the same for cellular immunofluorescence.
7
Quantitative Protein Expression Analysis
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PD-L1, PD-L2, ICAM-1, and IL-10 proteins were measured by Western blot analysis using anti-PD-L1 (Abcam, Cambridge, MA, USA), anti-PD-L2 (R&D systems, Minneapolis, MN, USA), anti-ICAM-1 (Cell Signaling Technology, Danvers, MA, USA), anti-IL-10 (Abcam), and anti-β-actin (Cell Signaling Technology) antibodies, respectively. Western blots were visualized using Super Signal West Pico Chemiluminescent Substrate (Thermo Fisher). The quantitative analysis of signal intensity was performed using Alpha View SA software (Protein simple, San Jose, CA, USA).
8
Curcumin-Collagen Hydrogel for Osteoarthritis
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Gelatin type B, tragacanth gum, Curcumin, Glacial acetic acid, Sodium sulfate, 37% formaldehyde, Chondroitin sulfate, EDC, NHS were purchased from Soleberg Reagent. Collagen (Type II, Chicken, Protein 60%, Yuanye), Fuchsin Complete Adjuvant (FCA), Fuchsin Incomplete Adjuvant (FICA) was purchased from Genye Reagent Company. Nitrendipine was obtained from Aladdin Chemical Reagent Co. CCK-8 kit, ROS kit, malondialdehyde (MDA) content assay kit, total glutathione (GSH-Px) content assay kit, superoxide dismutase (SOD) content assay kit and H&E kit were purchased from Beyotime Anti-HIF-1α, Anti-iNOS, anti-Arg-1, anti-IL-1β, anti-IL-10 were purchased from Abcam. RAW264.7 cells were obtained from Chinese Academy of Sciences.
9
Quantifying CD19 and IL-10 in Thymus Tissue
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The tissues slices were incubated with a polyclonal rabbit anti-IL-10 (bought from Abcam Company, Britain) and mouse monoclonal anti-CD19 (bought from Abcam Company, Britain) antibodies for 16 hours under 4°C at a dilution of 1:300 and 1:100, respectively. Subsequently, the slides were incubated with a mixture of secondary antibody for IF. Both secondary antibodies were used at a dilution of 1:100. Then, CD19/IL-10 Double immunofluorescence staining was observed under Confocal fluorescence microscopy (Olympus, Center Valley, PA, USA), and we performed semi-quantitative analysis by Software Tissuequest 4.0.1.0140. We stained the nucleus with DAPI, spot number of CD19 and IL-10 was collected by the software and DAPI spots number was seemed as reference. We calculated the spots number of CD19 and IL-10/spots number of DAPI, so as to obtain the relative cell counts of CD19 and IL-10 in thymus tissue of different groups.
10
Colon Tissue Histological Analysis
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Tissues from the mice or rats were freshly excised and fixed in 10% triformol. Once the samples were fixed, dehydration, clarification, and inclusion were carried out. After the blocks were obtained, the sections were cut using a microtome (Microm HM325, Germany), with a thickness of 5 μm. Sections of hydrated and deparaffinized tissues were stained with HE followed by appropriate method for histological observation. From each colon description, 10 sections were analyzed by three independent observers (JM, EM, and RMC).
The paraffin-embedded slices of colon tissue (4 μm) were incubated overnight with anti-NF-κB p65, anti-Foxp3, anti-IL-10, and anti-TNF-α primary antibodies at 4°C; all the antibodies were purchased from Abcam (Cambridge, UK). The slices were then washed with PBS and incubated with horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature. After washing with PBS again, the slices were developed using 3,3′-diaminobenzidine as a chromogen and counterstained with hematoxylin. Images were acquired using a Leica DM2500 system (Leica Microsystems, Germany).
The paraffin-embedded slices of colon tissue (4 μm) were incubated overnight with anti-NF-κB p65, anti-Foxp3, anti-IL-10, and anti-TNF-α primary antibodies at 4°C; all the antibodies were purchased from Abcam (Cambridge, UK). The slices were then washed with PBS and incubated with horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature. After washing with PBS again, the slices were developed using 3,3′-diaminobenzidine as a chromogen and counterstained with hematoxylin. Images were acquired using a Leica DM2500 system (Leica Microsystems, Germany).
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