Rhodamine 123, l ‐NAME, and sodium dodecyl sulfate (SDS) were purchased from Wako (Tokyo, Japan). ISOGEN was purchased from Nippongene (Tokyo, Japan). DMEM, RIPA buffer, protease inhibitor cocktail, and β‐Actin antibody were purchased from Sigma (MO, USA). Fetal bovine serum (FBS) and house serum (HS) were purchased from Gibco (NY, USA). MTT or 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide and l ‐NAME were purchased from Dojindo (Kumamoto, Japan). l ‐citrulline was supplied by Kyowa Hakko Bio Co., Ltd., Tokyo, Japan) while PGC‐1α (3G6) antibody was purchased from Cell Signaling Technology (Hertfordshire, UK). Glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) antibody (6C5) was purchased from Santa Cruz Biotechnology (CA, USA).
L name
Manufactured by Dojindo Laboratories
Sourced in Japan, United States
L-NAME is a chemical compound produced by Dojindo Laboratories. It functions as a synthetic inhibitor of nitric oxide synthase (NOS), the enzyme responsible for the production of nitric oxide in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti β actin antibody, by Santa Cruz Biotechnology (2 mentions)
Daf 2da, by Cayman Chemical (2 mentions)
Alexa fluor 555 anti rabbit, by Thermo Fisher Scientific (2 mentions)
H2dcfda, by Thermo Fisher Scientific (2 mentions)
Rabbit anti zo 1, by Thermo Fisher Scientific (2 mentions)
Sodium dodecyl sulfate (sds), by Fujifilm (1 mentions)
Rhodamine 123, by Fujifilm (1 mentions)
L name, by Fujifilm (1 mentions)
Glyceraldehyde 3 phosphate dehydrogenase gapdh antibody 6c5, by Santa Cruz Biotechnology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
β actin antibody, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Isogen, by Nippon Gene (1 mentions)
Csf1 m csf, by Thermo Fisher Scientific (1 mentions)
1h 1 2 4 oxadiazolo 4 3a quinoxalin 1 one odq, by Cayman Chemical (1 mentions)
Noc12, by Dojindo Laboratories (1 mentions)
Fetpps, by Cayman Chemical (1 mentions)
Amg9810, by Fujifilm (1 mentions)
8 protocols using l name
1
Analyzing Cellular Responses with Rhodamine 123 and L-NAME
2
Osteoclast Differentiation Protocol
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ddY mice were purchased from SHIMIZU Laboratory Supplies Co., Ltd. (Kyoto, Japan). Experiments were conducted according to the Animal Experiment Implementation Guidelines of Osaka Dental University (Approval number: 18-02014). Macrophage colony-stimulating factor 1 (CSF-1/M-CSF) and soluble NfϰB ligand (sRANKL) were purchased from PeproTech (Rocky Hill, NJ, USA). 1H-[1,2,4]oxadiazolo[4,3a]quinoxalin-1-one (ODQ) was purchased from Cayman Chemical (Ann Arbor, MI, USA), and l -NAME was purchased from Dojindo Molecular Technologies (Kumamoto, Japan). TH was synthesized by Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan).
3
Comprehensive Immunofluorescence Staining Protocol
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Rabbit anti-CLDN1, mouse anti-CLDN4, rabbit anti-ZO-1, Alexa Fluor 488 anti-mouse, and Alexa Fluor 555 anti-rabbit antibodies, and H2DCFDA were obtained from Thermo Fisher Scientific (San Diego, CA, USA). Goat anti-β-actin antibody was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse anti-phosphoserine (p-Ser) and anti-phosphothreonine (p-Thr) antibodies were from Sigma-Aldrich (Saint Louis, MO, USA). LY, NiSPY-3, Quest Fluo-8 AM, rabbit anti-nitrotyrosine, rabbit anti-NOS1, rabbit anti-NOS2, rabbit anti-NOS3, and rabbit anti-ubiquitine were from Biotium (Fremont, CA, USA), Goryo Kagaku (Hokkaido, Japan), AAT Bioquest (Sunnyvale, CA, USA), R&D Systems (Minneapolis, MN, USA), Cell Signaling Technology (Beverly, MA, USA), ProteinTech (Rosemont, IL, USA), GeneTex (Irvine, CA, USA), and Stressgen Biotechnologies Corporation (British Columbia, Canada), respectively. NOC12, SIN-1, and L-NAME were from Dojindo Laboratories (Kumamoto, Japan). AMG9810, olvanil, and RN1734 were from FUJIFILM Wako Pure Chemical Corporation (Tokyo, Japan). DAF-2DA and FeTPPs were from Cayman Chemical (Ann Arbor, MI, USA). All other reagents were of the highest grade of purity available.
4
Role of NO in Bile Duct Ligation
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To investigate the role of NO in BDL model, WT or Cygb−/− mice were administered L-NAME (Dojindo, MD, USA) in drinking water at the dose of 0.5 mg/ml for 9 days (n = 5 per each group). At day 7, the mice were performed BDL, and scarified 2 days later.
In NO donor treatment, WT or Cygb−/− mice were received 3 doses of SNP (2 mg/kg body weight) (Millipore Corp., Billerica, MA, USA) or saline by i.p. injection (n = 5 per each group). The first, second and third dose of SNP were injected immediately, 24 h, and 48 h, respectively, after the mice were subjected to BDL. Mice were scarified at 2 h after the last dose.
In NO donor treatment, WT or Cygb−/− mice were received 3 doses of SNP (2 mg/kg body weight) (Millipore Corp., Billerica, MA, USA) or saline by i.p. injection (n = 5 per each group). The first, second and third dose of SNP were injected immediately, 24 h, and 48 h, respectively, after the mice were subjected to BDL. Mice were scarified at 2 h after the last dose.
5
Evaluating Endothelial Cell Function
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Rabbit anti-CLDN1 and Alexa Fluor 555 anti-rabbit antibodies were obtained from Thermo Fisher Scientific (San Diego, CA, USA). Goat anti-β-actin antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). L-NAME, LY, NiSPY-3, Quest Fluo-8 AM, rabbit anti-nitrotyrosine, and rabbit anti-NOS3 antibodies were from Dojindo Laboratories (Kumamoto, Japan), Biotium (Fremont, CA, USA), Goryo Kagaku (Hokkaido, Japan), AAT Bioquest (Sunnyvale, CA, USA), R&D Systems (Minneapolis, MN, USA), and Cell Signaling Technology (Beverly, MA, USA), respectively. DAF-2DA, CAPE, apigenin, kaempferol, kaempferide, chrysin, quercetin, and apiin were from Cayman Chemical (Ann Arbor, MI, USA). EBGP was kindly provided from Yamada Bee Company (Okayama, Japan).
6
Evaluating Vasodilation Mechanisms
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7
Signaling Pathway Modulation Assay
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Reagent sources were as follows: MCT (Wako Pure Chemical Industries, Ltd., Osaka, Japan), l -NAME (Dojindo, Kumamoto, Japan), BAY11-7082 (Merck (Calbiochem), Darmstadt, Germany), PD98059 (Wako Pure Chemical Industries, Ltd.), SP600125 (Jena Bioscience, Jena, Germany) and SNP (Sigma Aldrich, St., Louis, MO, USA).
Antibody sources were as follows: anti-POSTN (1:100 dilution) (Proteintech, Rosemont, IL, USA), anti-glyceraldehyde-3-phosphate dehydrogenase (1:1000 dilution) (GeneTex, Irvine, CA, USA), anti-iNOS (1:250 dilution for western blotting or 1:100 dilution for immunohistochemistry) (Becton, Dickinson and Company, Franklin Lakes, NJ, USA or Bioss, Woburn, MA, USA), anti-total-actin (1:1000 dilution) (Sigma Aldrich), anti-phospho-VASP (1:500 dilution) (Abcam, Cambridge, UK), anti-phospho-ERK1/2 (1:1000 dilution), anti-phospho-JNK (1:250 dilution), anti-total-JNK (1:500 dilution), anti-phospho-NF-κB p65 (1:500 dilution) (Cell Signaling Technology, Madison, WI, USA), anti-total-ERK1/2 (1:100 or 1:200 dilution) (Santa Cruz Biotech, Santa Cruz, CA, USA or Bioss), anti-total-NF-κB p65 (1:500 dilution) (Santa Cruz Biotech), horseradish peroxidase (HRP)-conjugated anti-rabbit IgG and HRP-conjugated anti-mouse IgG (1:10,000 dilution) (Amersham Biosciences, Buckinghamshire, UK or Cell Signaling Technology).
Antibody sources were as follows: anti-POSTN (1:100 dilution) (Proteintech, Rosemont, IL, USA), anti-glyceraldehyde-3-phosphate dehydrogenase (1:1000 dilution) (GeneTex, Irvine, CA, USA), anti-iNOS (1:250 dilution for western blotting or 1:100 dilution for immunohistochemistry) (Becton, Dickinson and Company, Franklin Lakes, NJ, USA or Bioss, Woburn, MA, USA), anti-total-actin (1:1000 dilution) (Sigma Aldrich), anti-phospho-VASP (1:500 dilution) (Abcam, Cambridge, UK), anti-phospho-ERK1/2 (1:1000 dilution), anti-phospho-JNK (1:250 dilution), anti-total-JNK (1:500 dilution), anti-phospho-NF-κB p65 (1:500 dilution) (Cell Signaling Technology, Madison, WI, USA), anti-total-ERK1/2 (1:100 or 1:200 dilution) (Santa Cruz Biotech, Santa Cruz, CA, USA or Bioss), anti-total-NF-κB p65 (1:500 dilution) (Santa Cruz Biotech), horseradish peroxidase (HRP)-conjugated anti-rabbit IgG and HRP-conjugated anti-mouse IgG (1:10,000 dilution) (Amersham Biosciences, Buckinghamshire, UK or Cell Signaling Technology).
8
Inhibition of Nitric Oxide Synthase in Rats
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At ~24 h before LISW exposure, Group 3 rats were anesthetized with isoflurane inhalation (5% for induction; 1.5%-2.0% for maintenance) and then a small incision was made in the groin and a polyurethane catheter (MRE-040; ID, 1.02 mm; OD, 0.64 mm) was inserted into the femoral vein. The NO synthase inhibitor nitro-L-arginine methyl ester (L-NAME, Dojindo Laboratories, Kumamoto, Japan) diluted with saline at a concentration of 4.0 mg/mL and a volume of 5.0 mL/kg animal body weight was intravenously injected through the catheter. With this protocol, NO synthase activity in the brain was expected to be inhibited by ~50% (Iadecola et al., 1994) .
After injection, the femoral vein was ligated with a suture, and the incision was closed with a suture. Rats received L-NAME administration under isoflurane anesthesia as long as ~24 h before imaging, and the injection volume was within the standard value for laboratory animals (Turner et al., 2011) . Thus, we assumed that both the injection and anesthesia would not affect the results of imaging, and neither saline injection nor isoflurane anesthesia were applied to all groups of rats other than Group 3.
After injection, the femoral vein was ligated with a suture, and the incision was closed with a suture. Rats received L-NAME administration under isoflurane anesthesia as long as ~24 h before imaging, and the injection volume was within the standard value for laboratory animals (Turner et al., 2011) . Thus, we assumed that both the injection and anesthesia would not affect the results of imaging, and neither saline injection nor isoflurane anesthesia were applied to all groups of rats other than Group 3.
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