Peiminine was purchased from MedChem Express (Monmouth Junction, NJ, USA). ATP and Griess reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). The anti-phospho-NF-κB (p65) antibody was acquired from InvivoGen (San Diego, CA, USA). The anti-phospho-IκB, anti-IL-6, anti-MAPK, and anti-COX-2 antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-β-actin antibody was obtained from Santa Cruz Biotechnology (Dallas, TX, USA), and the anti-IL-1β antibody was obtained from R&D Systems (Minneapolis, MN, USA).
Anti il 6
Manufactured by Cell Signaling Technology
Sourced in United States, United Kingdom
Anti-IL-6 is a laboratory reagent that targets the cytokine Interleukin-6 (IL-6). It is used in research applications to detect and study the IL-6 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti il 1β, by Cell Signaling Technology (15 mentions)
Anti tnf α, by Cell Signaling Technology (10 mentions)
Anti β actin, by Cell Signaling Technology (9 mentions)
Anti nf κb p65, by Cell Signaling Technology (6 mentions)
Anti mcp 1, by Cell Signaling Technology (5 mentions)
Anti cox 2, by Cell Signaling Technology (4 mentions)
Anti stat3, by Cell Signaling Technology (4 mentions)
Anti p stat3, by Cell Signaling Technology (4 mentions)
Anti gapdh, by Cell Signaling Technology (4 mentions)
Ecl chemiluminescent detection system, by Cytiva (3 mentions)
Enhanced chemiluminescence system, by GE Healthcare (3 mentions)
Anti gapdh, by Proteintech (3 mentions)
Anti sirt1, by Cell Signaling Technology (3 mentions)
Anti nrf2, by Cell Signaling Technology (3 mentions)
Anti p erk1 2, by Cell Signaling Technology (2 mentions)
Anti c myc, by Cell Signaling Technology (2 mentions)
Anti akt, by Cell Signaling Technology (2 mentions)
Anti erk, by Cell Signaling Technology (2 mentions)
Anti p akt, by Cell Signaling Technology (2 mentions)
Anti p erk, by Cell Signaling Technology (2 mentions)
44 protocols using anti il 6
1
Peiminine Inhibits NF-κB Signaling
2
Immunohistochemical Analysis of Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry (IHC) analysis was carried out by using MaxVisionTM techniques (Maixin Bio, China) based on the manufacturer's instructions. Firstly, the lung tissue was fixed by 4% paraformaldehyde (Solarbio, Shanghai, China), dehydrated, and paraffin embedded. The 5 μm thick slides were obtained. The deparaffinization and hydration were performed, the slides were then incubated with 3% H2O2 (Sinopharm, China) for 10 min and 0.1% trypsin (Beyotime, China) for 20 min. The primary antibodies were incubated at 4°C overnight and then incubated with HRP-polymer-conjugated secondary antibodies at 37°C for 1 h. The slides were then stained by using the chromogenic reagent diaminobenzidine (DAB, Zhongshan, Beijing, China) for 3 min and counterstained with hematoxylin (Jiancheng, Nanjing, China). An inverted microscope (Olympus, Japan) was employed for image acquisition. The primary antibodies anti-IL-6, anti-JAK1, anti-STAT3, and anti-SOCS3 were purchased from Cell Signaling Technology (CST).
3
Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein samples were extracted using RIPA lysis buffer. A nucleic acid and protein microanalyzer (Molecular Devices, United States) was used to determine protein concentrations. Western blotting was performed according to a published protocol (Wu et al., 2022 (link)). Primary antibodies were incubated overnight at 4°C as follows: anti-iNOS (Proteintech, 22226-1-AP, 1:5000), anti-PD-L1 (Proteintech, 66248-1-Ig, 1:3000), anti-Arg-1 (Proteintech, 16001-1-AP, 1:2000), anti-GADPH (Proteintech, 60004-1-Ig, 1:10000), anti-CD9 (Proteintech, 20597-1-AP, 1:2000), anti-CD63 (Proteintech, 25682-1-AP, 1:2000), anti-TSG101 (Proteintech, 28283-1-AP, 1:3000), anti-JAK2 (Cell Signaling Technology, 3230S, 1:2000), anti-STAT3 (Proteintech, 10253-2-AP, 1:5000), anti-p-STAT3 (Cell Signaling Technology, 9145S, 1:1000), and anti-IL-6 (Cell Signaling Technology, 12912S, 1:1000). The HRP-conjugated secondary antibody (ZSGB-BIO, ZB-2305; ZB-2301) was incubated for 1 h at room temperature. The signal was detected using the ECL method (Beyotime, China, P0018FS). Band density was quantified using ImageJ software (NIH, Bethesda, MD, United States) for the grayscale intensity of each protein.
4
Immunohistochemical Analysis of Liver Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The entire left lobe of the mouse livers was excised and fixed in 4% paraformaldehyde for at least 24 h, embedded in paraffin, and cut to yield 5-µm-thick sections. Following hydration in a decreasing ethanol gradient, all sections were deparaffinized and stained with Harris hematoxylin solution for 5 min at 37°C. For IHC, paraffin-embedded liver tissue samples were cut into 5-µm-thick consecutive sections, dewaxed in xylene, and rehydrated in graded ethanol solutions. Then, the nonspecific binding sites in the tissues were blocked, and the steam cooking method was used for antigen retrieval. The sections were incubated with the following primary antibodies: Anti-p-STAT1 (1:50; product no. 7649; Cell Signaling Technology, Inc.); anti-IL-6 (1:100; cat. no. GB11117) and anti-IL-10 (1:100; cat. no. GB11108; both from Servicebio Technology Co., Ltd.) overnight at 4°C. Then, they were incubated with horseradish peroxidase-conjugated polyclonal goat anti-rabbit secondary antibodies (1:200; GB23303; Servicebio Technology Co., Ltd.) for 1 h at room temperature. Finally, the sections were counterstained with hematoxylin.
5
Detecting Cytokine Protein Levels
6
Evaluating Inflammatory Cytokines by Western Blot
7
IL11 and TGF-β1 Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant proteins: Mouse TGFβ1 (7666-MB, R&D Systems), mouse IL11 (rmIL11, UniProtKB: P47873, GenScript). Antibodies: anti-smooth muscle actin (ab7817, abcam), anti-Collagen I (ab34710, abcam), anti-IL11RA (ab125015, abcam), goat anti-mouse Alexa Flour 488-conjugated secondary antibody (ab150113, abcam), goat anti-rabbit Alexa Flour 488-conjugated secondary antibody (ab150077, abcam), DAPI (D1306, Thermo Fisher Scientific). Primary antibodies for Western blots include: anti-Fibronectin antibody (ab2413, Abcam); anti-IL6 (12912, Cell Signaling); anti-p-ERK1/2 (4370, Cell Signaling), anti-ERK1/2s (4695, Cell Signaling), anti-p-SMAD2 (ab216482, Abcam), anti-p-NF-kB (ab28856, Abcam), anti-p-STAT3 (4113, Cell Signaling), anti-SMAD2 (5339, Cell Signaling), anti-NF-kB (8242, Cell Signaling), anti-STAT3 (4904, Cell Signaling) and anti-GAPDH (2118, Cell Signaling). Monoclonal anti-IL11 antibody (X203), generated in our previous study15 (link), was used to detect IL11 protein expression in tissue lysates. Secondary antibodies for Western blots include: anti-rabbit HRP (7074, Cell Signaling) or anti-mouse HRP (7076, Cell Signaling).
8
Protein Expression Analysis in Cell Lysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed on ice for 30 min in a buffer containing 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Nonidet P-40, and 0.1% SDS supplemented with protease inhibitors (10 g/ml leupeptin, 10 g/ml pepstatin A, and 10 g/ml aprotinin). The proteins were separated by SDS-PAGE, transferred to a nitrocellulose membrane, and detected using anti-IL-6 (#12912, Cell Signaling Technology, Danvers, MA, USA), anti-p-STAT3 (Y705) (#9145, CST), anti-STAT3 (#9139, CST), anti-non-phospho (active) β-catenin (#19807, CST), and anti-β-actin (#3700, CST) antibodies. The proteins were visualized using an enhanced chemiluminescence system (GE Healthcare, Piscataway, NJ, USA).
9
Kidney Cell Protein Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kidney and HK‐2 cells protein were extracted using RIPA buffer including protease inhibitor and PMSF (Heart, China). Samples protein concentration was measured by BCA Protein Quantitative Kit (Heart, China) and then adjusted to equal amounts for running western blot. Antibodies: anti‐p16 (1:1000; Abcam, England), anti‐p21 (1:1000; Abcam, England), anti‐p53 (1:1000; Abcam, England), anti‐Klotho (1:1000; Abcam, England), anti‐NLRP3 (1:1000; Cell Signaling Technology, USA), anti‐eNOS (1:1000; Cell Signaling Technology, USA), anti‐TGF‐β (1:1000; Abcam, England), anti‐IL‐6 (1:1000; Cell Signaling Technology, USA), anti‐IL‐1β (1:1000; Cell Signaling Technology, USA), anti‐Smad3 (1:1000; Proteintech, China), anti‐LTBP2 (1:1000; Santa cruz, USA), anti‐Tubulin (1:3000; Proteintech, China) and anti‐Gapdh (1:3000; Proteintech, China) were applied. Membranes were detected by chemiluminescence gel imaging system (FluorChem E; ProteinSimple , USA). Quantification of protein abundance was performed by Image J software 1.8.0.
10
Molecular Mechanisms of DHED Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
17β-dihydroxyestra-1,4-diene-3-one (DHED) and ICI182780 were purchased from Sigma-Aldrich (St. Louis, MO) and Selleck (TX, USA), respectively. The primary antibodies used for Western blot analysis were as follows: anti-TNF-α, anti-IL1β, anti-IL-6, anti-klf5, anti-p-IκB kinase (IKK), anti-inhibitor of NF-κB α (IκBα), anti-tau, anti-p-tau (ser235), and anti-p-tau (ser396) were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-GAPDH was from Proteintech Group (Wuhan, China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!