PF [C23H28O11, MW: 480.45, purity: 98.78% (HPLC), LD50: 9,530 mg/kg, Figure 1A ] was obtained from Nanjing GOREN BIO Technology Co., Ltd. (Nanjing, China). PF was dissolved in saline to provide a stock solution. A microalbumin assay kit was acquired from Abcam Biotechnology (Abcam, Cambridge, United Kingdom). An immunohistochemistry kit (PV-9000) was purchased from Beijing Zhongshan Biotechnology Inc. (Zhongshan, China). Rabbit anti-TLR4, anti-MyD88, anti-Trif, and anti-iNOS antibodies were purchased from Abcam Biotechnology (Abcam, Cambridge, United Kingdom) and anti-p-IRF3, anti-NF-κB p65, and anti-NF-κB p-p65 antibodies were obtained from Cell Signaling Technology (CST Beverly, MA, United States). Rabbit anti-p-IRAK1 and anti-CD68 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, United States). Other materials for Western blotting (WB) were acquired from Amersham Life Science (Little Chalfont, United Kingdom). TRIzol was obtained from Invitrogen (Invitrogen, CA, United States), and the SYBR Green PCR Master Mix Kit was purchased from Bio-Rad (Bio-Rad, CA, United States). A cDNA synthesis kit was purchased from Promega (Promega, Madison, WI, United States). GAPDH and TNF-α primers were obtained from Shanghai Sangon Company (Shanghai, China).
Anti inos antibody
Manufactured by Abcam
Sourced in United States, United Kingdom, Germany
Anti-iNOS antibody is a laboratory reagent used for the detection and quantification of inducible nitric oxide synthase (iNOS) protein in various biological samples. iNOS is an enzyme responsible for the production of nitric oxide, a signaling molecule involved in various physiological and pathological processes. This antibody can be used in techniques such as Western blotting, immunohistochemistry, and ELISA to study the expression and localization of iNOS in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Anti il 1β antibody, by Abcam (4 mentions)
Anti nf κb p65, by Cell Signaling Technology (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Anti f4 80 antibody, by Abcam (2 mentions)
Anti cd11b antibody, by Abcam (2 mentions)
Anti arg 1 antibody, by Abcam (2 mentions)
Anti cleaved caspase 3 antibody, by Cell Signaling Technology (2 mentions)
Anti neun antibody, by Abcam (2 mentions)
Rabbit anti oxr1, by Merck Group (2 mentions)
Anti gpx1 antibody, by Abcam (2 mentions)
Anti cd3, by Abcam (2 mentions)
Anti iba1, by Abcam (2 mentions)
Anti ki67 antibody, by BD (2 mentions)
Anti γh2ax antibody, by Cell Signaling Technology (2 mentions)
F4 80 antibody, by Abcam (2 mentions)
Anti phospho stat3 tyr705, by Cell Signaling Technology (2 mentions)
Goat anti rat, by Thermo Fisher Scientific (2 mentions)
Anti epcam antibody, by Santa Cruz Biotechnology (2 mentions)
Anti nitrotyrosine antibody, by Merck Group (2 mentions)
Anti mpo antibody, by Abcam (2 mentions)
35 protocols using anti inos antibody
1
TLR4-Mediated Inflammatory Response Modulation
2
Macrophage Phenotype Analysis in 3D Scaffold
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For phenotype analysis, the macrophages were co-cultured with the sterilized immuno-functioned 3D-scaffolds placed in transwells for 3 days. After fixation and permeabilization, the rBMDMs were stained with anti-iNOS antibodies (Abcam), anti-CD68 antibodies (Abcam), and DAPI, then observed on the confocal microscope to determine M1 phenotypic ratio. With the same process, rBMDMs were stained with anti-CD206 antibodies, anti-CD68 antibodies, and DAPI to determine the M2 phenotypic ratio. Immunofluorescence images were obtained on a confocal microscope, and the ratios of CD206+ macrophage and iNOS+ macrophages were analyzed.
3
Wound Healing Evaluation in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At day 7 and 14 after injury, the mice were euthanized, and skin samples were harvested and fixed. Hematoxylin-Eosin (H&E) and Masson's staining were performed to detect wound healing and collagen deposition at the injured sites. Immunofluorescence staining was carried out to determine the angiogenic and macrophage effects at different time points. To detect angiogenic effects, rat anti-mouse CD31 (Abcam, Cambridge, MA) and Alexa Fluor 594 goat anti-rat IgG (Invitrogen, Grand Island, NY) were used. To track macrophages, anti-F4/80 (Abcam, Cambridge, UK), anti-CD68 (Abcam), anti-RELM-α (Abcam), anti-CD206 (Abcam), and anti-iNOS antibodies (Abcam) were used. Moreover, anti-α-SMA antibody (Boster Bio-Engineering Company, Wuhan, China) was used to evaluate infiltration of myofibroblasts at the injured sites. Alexa Fluor 488 and 594 (Invitrogen, Carlsbad, CA) were applied appropriately. The cell nuclei were counter-stained with 4', 6-diamidino-2-phenylindole (DAPI). Immunohistochemistry was performed to detect the expression of interleukin-1β (IL-1β) (Boster Bio-Engineering Company, Wuhan, China). The images were analyzed by ImageJ software.
4
Immunohistochemical Analysis of Cardiac Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical staining of left ventricle specimens was performed as we recently reported [20 (link)]. Antigen retrieval was carried out by boiling the sections in 10 mM citrate buffer (pH 6), followed by incubation with anti-iNOS antibodies (Abcam, cat # ab15323, Cambridge, UK) and anti-desmin antibodies (Cat # ab131442, Abcam, Cambridge, UK). The sections were incubated with secondary antibodies for 30 min at room temperature. The sections were counter stained with Meyer’s hematoxylin.
Morphometry of the areas% of collagen deposition and the areas% of iNOS and desmin immunostaining was performed using a “Leica Qwin 500 C” image analyzer (Cambridge, UK) in 10 non-overlapping fields for each group. Quantitative data were summarized as means and standard deviations (SD) and compared using analysis of variance (ANOVA) followed by post-hoc analysis (Tukey test). A p-value < 0.05 was considered statistically significant. Calculations were carried out using a statistical package of social science (SPSS) software, version 19.
Morphometry of the areas% of collagen deposition and the areas% of iNOS and desmin immunostaining was performed using a “Leica Qwin 500 C” image analyzer (Cambridge, UK) in 10 non-overlapping fields for each group. Quantitative data were summarized as means and standard deviations (SD) and compared using analysis of variance (ANOVA) followed by post-hoc analysis (Tukey test). A p-value < 0.05 was considered statistically significant. Calculations were carried out using a statistical package of social science (SPSS) software, version 19.
5
Wogonoside Modulates Inflammatory Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wogonoside (Wog) was acquired from Biopurify Phytochemicals Ltd. (Sichuan, China). LPS (0111:B4) was acquired from Sigma-Aldrich (Shanghai, China). Griess reagent was supplied by Promega Biotech Co (Madison, WI, USA), and TRIzol reagent was acquired from Invitrogen Co. (Carlsbad, CA, USA). Dichlorofluorescein diacetate (DCFH-DA) was obtained from Solarbio Technology Inc. (Beijing, China). RT SuperMix and Universal SYBR qPCR Master were both supplied by Vazyme Biotech (Nanjing, China). SP600125, brefeldin A, and CCK-8 were obtained from Selleck Chemicals (Houston, USA). An ECL kit and PVDF membrane were purchased from Millipore Company (Shanghai, China). The anti-COX-2 antibody was purchased from Proteintech (Wuhan, China). The anti-IL-1β, anti-phospho-p65, anti-TNF-α, anti-p65, anti-IL-6 antibodies, anti-phospho-MAPK family, and anti-MAPK family antibody samplers were supplied by Cell Signaling Technology, Inc. (Boston, MA, USA). HRP-goat anti-rabbit/mouse IgG antibody, anti-c-Jun, and anti-iNOS antibodies were obtained from Abcam (Cambridge, MA, USA). GAPDH antibody was acquired from TransGen Biotech (Beijing, China). Alexa Fluor 488-labeled and 594-labeled secondary antibodies were purchased from Invitrogen (Shanghai, China).
6
Characterizing Macrophage Activation and Inflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Main instruments: Vernier caliper (Meinaite, Germany), Ultrasound imaging equipment (Vevo 3100), dehydrator, embedding machine, slicer, spreader (Leica, Germany), automatic tissue scanner (VS120, Olympus).
Main reagents: Cynarin (Chengdu Pufei De Biotechnology, 17041106), xylene (Sinopharm, 10023418), Ethylenediamine tetraacetic acid (Sinopharm. 10009617), 75%Ethanol (Sinopharm, 80176961), Absolute ethanol (Sinopharm, 10009218), Hematoxylin(sigma, H3136), Eosin Y (sigma, E4009), Monosodium Urate (sigma, U2875), Bovine Serum Albumin (Aladdin, B265994),Anti-F4/80 antibody (Abcam, ab6640), Macrophage-stimulating factor (MCSF) (Peprotech, 315–02), Anti-iNOS antibody (Abcam, ab3523), VECTASHIELD Mounting Medium with DAPI (Vectorlabs, H-1200), Trypsin antigen repair solution (Leagene Biotechnology, IH0310), Cell Counting Kit-8 (Dojindo laboratories, CK04), PrimeScript™ RT Reagent Kit (TaKaRa, RR037A), qPCR SYBR Green Master Mix (YESEN, 11201ES03), p-p65 antibody (CST, 3033), p65 antibody (CST, 8242), GAPDH antibody (sigma, G9545), p-p38 antibody (CST, 4511), p-IKKa/β antibody (CST, 2697),Anti-rabbit antibody (CST, 7074), p-JNK antibody (CST, 4668), p-ERK1/2 antibody (CST, 4370), Caspase 1/p20/p10 antibody (Proteintech, 22915-1-AP), Anti-NLRP3 antibody (Abcam, ab263899), Anti-IL-1β antibody (Abcam, ab200478).
Main reagents: Cynarin (Chengdu Pufei De Biotechnology, 17041106), xylene (Sinopharm, 10023418), Ethylenediamine tetraacetic acid (Sinopharm. 10009617), 75%Ethanol (Sinopharm, 80176961), Absolute ethanol (Sinopharm, 10009218), Hematoxylin(sigma, H3136), Eosin Y (sigma, E4009), Monosodium Urate (sigma, U2875), Bovine Serum Albumin (Aladdin, B265994),Anti-F4/80 antibody (Abcam, ab6640), Macrophage-stimulating factor (MCSF) (Peprotech, 315–02), Anti-iNOS antibody (Abcam, ab3523), VECTASHIELD Mounting Medium with DAPI (Vectorlabs, H-1200), Trypsin antigen repair solution (Leagene Biotechnology, IH0310), Cell Counting Kit-8 (Dojindo laboratories, CK04), PrimeScript™ RT Reagent Kit (TaKaRa, RR037A), qPCR SYBR Green Master Mix (YESEN, 11201ES03), p-p65 antibody (CST, 3033), p65 antibody (CST, 8242), GAPDH antibody (sigma, G9545), p-p38 antibody (CST, 4511), p-IKKa/β antibody (CST, 2697),Anti-rabbit antibody (CST, 7074), p-JNK antibody (CST, 4668), p-ERK1/2 antibody (CST, 4370), Caspase 1/p20/p10 antibody (Proteintech, 22915-1-AP), Anti-NLRP3 antibody (Abcam, ab263899), Anti-IL-1β antibody (Abcam, ab200478).
7
Immunofluorescence Staining Procedure
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed with PBS and fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.2% Triton X-100 for 20 min, and blocked with PBS containing 3% bovine serum albumin (BSA, Yeasen, China) for 1 h. Then, cells were incubated overnight with the following primary antibodies: anti-F4/80 antibody (eBioscience, USA), anti-iNOS antibody (Abcam, USA), anti-ARG1 antibody (Proteintech, China), anti-Collagen X antibody (Abcam), USA), and anti-Collagen II antibody (Abcam, USA). Alexa 488 or Alexa 594 dye-labeled secondary antibodies (Jackson, USA) were added and incubated for 1 h at room temperature. Nuclear staining was performed for 5 min by the addition of 4',6-diamidino-2-phenylindole (Beyotime, China) solution. Immunoreactivity was visualized using a fluorescence microscope (Carl Zeiss, Germany).
8
Immunofluorescent Staining of Macrophage Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After deparaffinization and rehydration through standard protocols, the paraffin sections were washed by PBS. Then sections were alternately bathed in boiling sodium citrate buffer for 20 min. After returning to room temperature, sections were washed. The membrane was permeated in 0.3% PBST (100 ml PBS, 0.3 ml Triton X-100) for 20 min. Sections were washed and then blocked by donkey serum at 37°C for 30 min. Sections were incubated by primary antibodies overnight at 4°C, including anti-F4/80 antibody (1:100, abcam), anti-INOS antibody (1/100, abcam), and anti-CD206 antibody (1:100, R&D). After incubtion the sections were restored to room temperature, washed by PBS, and then incubated by secondary antibodies, including donkey secondary antibodies to rat (1:2,000, Alexa Fluor®488, abcam), donkey secondary antibodies to rabbit (1:2,000, Alexa Fluor®647, abcam), and donkey secondary antibodies to goat (1:2,000, Alexa Fluor®555, abcam). After antibody incubation, sections were washed and finally sealed in a sealant containing DAPI. Images were acquired through a Leica confocal laser scanning microscopy (Leica).
9
Immunofluorescence Analysis of TLR4 and iNOS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells grown on the bottom of dishes were treated with PF for half an hour, exposed to HG levels for 24 h, and fixed with 4% paraformaldehyde for 15 min. Cell samples were blocked with 2% donkey serum albumin (EMD Millipore Corporation, United States) and stained with an anti-TLR4 antibody (Santa Cruz Biotechnology, CA, United States, 1:50) and anti-iNOS antibody (Abcam Biotechnology, Cambridge, United Kingdom, 1:50), followed by FITC-conjugated IgG (Santa Cruz Biotechnology, CA, United States, 1:200) and PE-conjugated IgG (Santa Cruz Biotechnology, CA, United States, 1:200). The nucleus was counterstained with 0.25 mg/ml 4-,6-diamidino-2-phenylindole (DAPI, Beyotime Institute of Biotechnology Jiangsu, China) for 5 min, the sections were mounted with antifade fluorescence mounting medium, and the macrophages were visualized using a Leica TCS SP5 laser confocal microscope (Leica, Germany) with the appropriate filters.
10
Western Blot Analysis of Autophagy Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot was performed by using a standard protocol as previously described [29 (link)]. Samples were normalized to 1 mg/mL and the loading volume was 20 mL/well. The membranes were, respectively, incubated with rabbit polyclonal anti-LC-3B (1 : 500; Novus Biologicals), rabbit polyclonal Beclin-1 (1 : 1000; Abcam Cambridge, UK), rabbit polyclonal ZIP8 (1 : 1000; Santa Cruz Biotechnology), rabbit polyclonal nuclear factor-kappa beta (NF-κβ) (1 : 1000, Abcam Cambridge, UK), rabbit polyclonal anti-iNOS antibody (1 : 1000, Abcam Cambridge, UK), and rabbit anti-β-actin (1 : 500; Santa Cruz Biotechnology). The bound antibodies were detected using goat anti-rabbit IgG-HRP antibody (1 : 1000; Abcam Cambridge, UK). The protein bands were visualized by an ECL detection system (Pierce Chemical, Rockford, IL, USA) and quantified by Image J software (NIH, Bethesda, MD).
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!