Liver tissues were homogenized in protein lysis buffer containing protease inhibitor. The protein concentration was determined by using bovine serum albumin as the standard. Proteins were denatured with 5× loading buffer in a heating block for 5 minutes. Equal amounts of whole liver tissue protein extracts were analyzed by SDS-polyacrylamide gel electrophoresis and standard western blotting analysis using antitumor necrosis factor (TNF)-α, anti-interferon (IFN)-γ, anti-inducible nitric oxide synthase (iNOS), and anti-NR1H4 antibodies (1:500, respectively; Abcam, Cambridge, MA, USA). The level of each protein was normalized to that of the housekeeping gene β-actin (1:2000; PPLYGEN, Beijing, China) or GAPDH (1:5000; Abcam) in the same sample using Gel-pro software (Media Cybernetics, LP, MD, USA).
Anti inducible nitric oxide synthase
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-inducible nitric oxide synthase (iNOS) is a laboratory product that detects the expression of the inducible nitric oxide synthase enzyme. iNOS is an enzyme involved in the production of nitric oxide, a signaling molecule with important physiological functions. This product can be used to analyze the levels and distribution of iNOS in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd68, by Abcam (3 mentions)
Gapdh, by Abcam (1 mentions)
Anti interferon ifn γ, by Abcam (1 mentions)
Gel pro software, by Media Cybernetics (1 mentions)
Tnf α, by Abcam (1 mentions)
Ammonium bicarbonate, by Thermo Fisher Scientific (1 mentions)
Docosahexaenoic acid dha, by Cayman Chemical (1 mentions)
Ezq protein assay kit, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Modified porcine trypsin, by Promega (1 mentions)
Prostaglandin e2 pge2 elisa kit, by Cayman Chemical (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Restore plus western blot stripping buffer, by Thermo Fisher Scientific (1 mentions)
Wst 1 assay kit, by Takara Bio (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Monoclonal anti β actin peroxidase, by Merck Group (1 mentions)
Chemidoc, by Bio-Rad (1 mentions)
Anti cyclooxygenase 2 cox 2, by Abcam (1 mentions)
Lysis buffer, by Merck Group (1 mentions)
8 protocols using anti inducible nitric oxide synthase
1
Hepatic Protein Expression Analysis
2
Proteomic Analysis of DHA and PGE2 Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Docosahexaenoic
acid (DHA, 100 mg in 0.4 mL of ethanol) and prostaglandin E2 (PGE2)
ELISA kit (#514010) were purchased from Cayman Chemical (Ann Arbor,
MI). Dithiothreitol (DTT), iodoacetamide (IAA), fetal bovine albumin
(FBS), formic acid (FA, mass spectrometry grade), and monoclonal anti-β-actin
peroxidase (#A3854) were purchased from Sigma-Aldrich (St. Louis,
MO). The WST-1 assay kit was obtained from Clontech (Mountain View,
CA). Dulbecco’s modified Eagle’s medium (DMEM) and penicillin/streptomycin
were obtained from Life Technologies (Grand Island, NY). Antibodies
of phospho-NF-κB p65 (Rabbit mAb #3033) and NF-κB p65
(Rabbit mAb #8242) were from Cell Signaling (Beverly, MA). Anti-inducible
nitric oxide synthase (iNOS) (#ab15323) was from Abcam (Cambridge,
MA). Urea (electrophoresis grade), thiourea (electrophoresis grade),
sodium dodecyl sulfate (SDS), ammonium bicarbonate, BCA protein assay
kit, EZQ protein assay kit, SuperSignal west pico plus chemiluminescent
substrate, Restore PLUS Western blot stripping buffer, tumor necrosis
factor α (TNF-α) mouse uncoated ELISA kit (#88-7324-88),
C18 tips, dimethyl sulfoxide (DMSO), water (H2O, HPLC grade), and acetonitrile (ACN, HPLC grade) were obtained
from Thermo Fisher (Waltham, MA). Modified porcine trypsin was acquired
from Promega (Madison, WI).
acid (DHA, 100 mg in 0.4 mL of ethanol) and prostaglandin E2 (PGE2)
ELISA kit (#514010) were purchased from Cayman Chemical (Ann Arbor,
MI). Dithiothreitol (DTT), iodoacetamide (IAA), fetal bovine albumin
(FBS), formic acid (FA, mass spectrometry grade), and monoclonal anti-β-actin
peroxidase (#A3854) were purchased from Sigma-Aldrich (St. Louis,
MO). The WST-1 assay kit was obtained from Clontech (Mountain View,
CA). Dulbecco’s modified Eagle’s medium (DMEM) and penicillin/streptomycin
were obtained from Life Technologies (Grand Island, NY). Antibodies
of phospho-NF-κB p65 (Rabbit mAb #3033) and NF-κB p65
(Rabbit mAb #8242) were from Cell Signaling (Beverly, MA). Anti-inducible
nitric oxide synthase (iNOS) (#ab15323) was from Abcam (Cambridge,
MA). Urea (electrophoresis grade), thiourea (electrophoresis grade),
sodium dodecyl sulfate (SDS), ammonium bicarbonate, BCA protein assay
kit, EZQ protein assay kit, SuperSignal west pico plus chemiluminescent
substrate, Restore PLUS Western blot stripping buffer, tumor necrosis
factor α (TNF-α) mouse uncoated ELISA kit (#88-7324-88),
C18 tips, dimethyl sulfoxide (DMSO), water (H2O, HPLC grade), and acetonitrile (ACN, HPLC grade) were obtained
from Thermo Fisher (Waltham, MA). Modified porcine trypsin was acquired
from Promega (Madison, WI).
3
Apoptotic Protein Expression in Intestine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The frozen intestine tissue was homogenized (1:9, w/v) with Lysis buffer (Sigma-Aldrich) containing a protease inhibitor cocktail (Sigma-Aldrich) and a phosphatase inhibitor cocktail (Sigma-Aldrich), and centrifuged at 12,000× g at 4 °C for 20 min to isolate the cellular proteins in the supernatant. To investigate protein expression related to apoptotic changes, we performed Western blotting according to a previous study [47 (link)]. After blocking with bovine serum albumin blocking buffer, membranes were incubated with various primary antibodies, such as anti-Annexin A1 (Anex1, 1:1000, Abcam, Cambridge, MA, USA), anti-formylpeptide receptor-like 1 (ALX/FPRL-1, 1:1000, Abcam), anti-inducible nitric oxide synthase (iNOS, 1:1000, Abcam), anti-cyclooxygenase 2 (COX-2, 1:1000, Abcam), anti-TNF-α (1:1000, Abcam), anti-proliferating cell nuclear antigen (PCNA; 1:1000, Abcam), anti-CD44 (1:1000, Abcam), and β-actin (1:1000, Abcam). After washing and adjusting the secondary antibody according to the manufacturer’s protocol, quantitative analysis of each protein band was performed using ChemiDoc (Bio-Rad Laboratories, Hercules, CA, USA).
4
Immune Regulation by PMA, Ionomycin, and OVA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phorbol 12-myristate 13-acetate (PMA; #P1585), ionomycin (Iono; #I0634), and OVA (#A5503) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Aluminum hydroxide (alum, #77161) was purchased from Thermo Fisher Scientific (Waltham, MA, USA) and C20 (#S6577) from Selleckchem (Houston, TX, USA). Antibodies against phospho(p)-PKCθ (#9377), p-STAT6 (#5654), STAT6 (#5397), p-NF-κB (#3033), and NFAT (#4389) were obtained from Cell Signaling Technology (Danvers, MA, USA), and anti-PKCθ (#sc-212), anti-β-actin (#sc-47778), and anti-lamin B1 (#sc-374015) antibodies from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-GATA3 (#ab61052). Anti-inducible nitric oxide synthase (iNOS; #ab136918) antibodies were purchased from Abcam (Cambridge, UK).
5
Multimarker Immunostaining of Kidney Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After being dehydrated with 30% sucrose and embedded in optimal cutting temperature compound (OCT), the kidney tissue was sliced by a freezing microtome at a thickness of 7 μm and stained with the following fluorescent-conjugated antibodies: anti-CD34 (Abcam, Shanghai, China), anti-CD105 (Abcam, Shanghai, China), anti-CD68 (Abcam, Shanghai, China), anti-inducible nitric oxide synthase (iNOS) (Abcam, Shanghai, China), anti-CD206 (Abcam, Shanghai, China), anti-VEGFA (Abcam, Shanghai, China), and Fluor 555conjugated α-SMA (Abcam, Shanghai, China). Alexa Fluor 405-, Alexa Fluor 488-or Alexa Fluor 555conjugated secondary antibodies against rat or rabbits were used to visualize antigen-antibody complexes. After staining, the sections were incubated with 4',6-diamidino-2-phenylindole (DAPI) for nuclear staining and sealed for photography using a Leica SP8 confocal microscope (Leica, Wetzlar, Germany).
6
Quantifying Macrophage Polarization in Femoral Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Femoral sections were prepared as previously described.40 (link),41 (link) Briefly, the femora were harvested, fixed overnight in 4% paraformaldehyde, demineralized in 0.5 M ethylenediaminetetraacetic acid (EDTA, pH 7.4), and embedded in optimal cutting temperature compounds. The region of interest (ROI) located 3 mm from the distal end of the femur was cut into 10 μm-thick transverse sections. Immunofluorescent staining for macrophage was performed to confirm the inflammatory conditions. Macrophages were identified by anti-CD11b antibody (1:500; Abcam, USA), M1 proinflammatory macrophages by anti-inducible nitric oxide synthase (iNOS) (1:500; Abcam), and M2 anti-inflammatory macrophages by anti-liver Arginase 1 (Arg1) antibody (1:500; Abcam). The iNOS/CD11b and the Arg1/CD11b double-positive cells were calculated as the M1 and M2 macrophages, respectively, using QuPath (GNU General Public License v3.0).42 (link)
7
Ghrelin Modulation of Neuroinflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ghrelin was obtained from Enzo Life Sciences (New York, USA) and dissolved in PBS. A bicinchoninic acid (BCA) protein assay kit was obtained from Beyotime Biotechnology (Shanghai, China). Nitrous oxide (NO) and lactate dehydrogenase (LDH) assay kits were purchased from the Nanjing Jiancheng Institute of Biological Engineering (Nanjing, China). Anti-NLRP3 and anti-apoptosis-associated speck-like protein containing a CARD (ASC) antibodies were obtained from Novus Biologicals (Centennial, USA). Anti-caspase-1, anti-CD68, and anti-inducible nitric oxide synthase (iNOS) antibodies were obtained from Abcam (Cambridge, UK), anti-IL-1β, anti-NF-κB P65, and anti-NF-κB p-P65 antibodies were obtained from Cell Signaling Technology (Danvers, USA). An anti-GSDMD antibody was obtained from Santa Cruz Biotechnology (Texas, USA). Anti-iba1 and anti-β-actin antibodies were obtained from Proteintech (Chicago, IL, USA). The luxol fast blue (LFB) stain fluid was obtained from Sigma-Aldrich (St. Louis, MO, USA). The enzyme-linked immunosorbent assay (ELISA) kits for rat IL-1β, IL-6, and tumor necrosis factor-α (TNF-α) were purchased from Dakewe Biotech (Shenzhen, China). The ELISA kit for rat IL-18 was purchased from RayBiotech (Norcross, USA).
8
Protein Expression Profiling in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed with lysis buffer containing a protease/phosphatase inhibitor cocktail (Cell Signaling Technology, Danvers, MA, USA). We used anti-S100A9 (Abcam for humans and R&D for mice), anti-phospho-S100A9 (ThermoFisher Scientific), anti-phospho-ERK (Abcam), anti-ERK (Abcam), anti-phospho-p38 (Abcam), anti-p38 (Abcam), anti-phospho-NF-κB (Abcam), anti-NF-κB (Cell Signaling Technology), anti-phospho‐Smad 3 (Abcam), anti-Smad 3 (Abcam), anti-MPO (Abcam), anti-NE (Santa Cruz Biotechnology), anti-CD68 (Abcam), anti-inducible nitric oxide synthase (iNOS) (Abcam), and anti-Arginase 1 (ThermoFisher Scientific) antibodies.
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!