Luteolin (≥ 98%, HPLC) was purchased from Sigma-Aldrich (Shanghai, China) and dissolved in DMSO as a stock solution. Enzyme-linked immunosorbent assay (ELISA) kits for human and mouse IFN-α and IFN-β were purchased from R&D Systems (Minneapolis, MN). Antibodies against p-STAT1Tyr701, STAT1, p-JAK1(Tyr1034/1035), JAK1, SOCS1, SOCS2, SOCS3, SOCS6, SHP-1, SHP-2 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against CIS and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). A Cell Counting Kit-8 (CCK-8) and horseradish peroxidase-conjugated goat anti-rabbit antibody were purchased from Beyotime Institute of Biotechnology (Haimen, China).
Horseradish peroxidase conjugated goat anti rabbit antibody
Manufactured by Beyotime
Sourced in China
Horseradish peroxidase-conjugated goat anti-rabbit antibody is a secondary antibody conjugated with the enzyme horseradish peroxidase. It is used to detect and quantify the presence of primary rabbit antibodies in various immunoassays and immunochemical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Reactive oxygen species assay kit, by Beyotime (2 mentions)
Cisplatin, by Merck Group (2 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Luteolin, by Merck Group (1 mentions)
Socs3, by Cell Signaling Technology (1 mentions)
Shp 1, by Cell Signaling Technology (1 mentions)
Antibodies against p stat1tyr701, by Cell Signaling Technology (1 mentions)
N acetylcysteine nac, by MedChemExpress (1 mentions)
Apoptosis detection kit, by BD (1 mentions)
Phospho nf κb pp 65, by Cell Signaling Technology (1 mentions)
Cck 8 assay kit, by Apexbio (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hela cells, by National Collection of Authenticated Cell Cultures (1 mentions)
Bhk 21, by Thermo Fisher Scientific (1 mentions)
Llc pk1 cells, by American Type Culture Collection (1 mentions)
Alexa fluor 594 conjugated donkey anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 488 conjugated donkey anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase conjugated goat anti mouse antibody, by Beyotime (1 mentions)
Bhk 21 cells, by National Collection of Authenticated Cell Cultures (1 mentions)
7 protocols using horseradish peroxidase conjugated goat anti rabbit antibody
1
Luteolin Modulates Interferon Signaling
2
Molecular Mechanisms of ISL-Sensitized Cisplatin
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ISL was purchased from Selleck Chemicals (Shanghai, China), and cisplatin was obtained from Sigma-Aldrich (St. Louis, MO, United States). Antibodies against cleaved caspase-3 and caspase-3 (19677-1-AP), Bax (50599-2-Ig), Bcl-2 (26593-1-AP), β-actin and horseradish peroxidase-conjugated goat anti-mouse antibody (SA00001-1) were purchased from Proteintech Group (Rosemont, IL, United States). Antibodies against SOD2, NF-κB (p-65) and Phospho-NF-κB (Pp-65) were purchased from Cell Signaling Technology (Beverly, MA, United States). Horseradish peroxidase-conjugated goat anti-rabbit antibody and the reactive oxygen species (ROS) assay kit were purchased from Beyotime Biotech (Nantong, China). An apoptosis detection kit was purchased from BD Biosciences (San Diego, CA, United States). The CCK-8 assay kit was purchased from ApexBio (MA, United States). N-Acetylcysteine (NAC) was purchased from MedChemExpress. All other reagents were of reagent grade or better, and deionized water was used for all the experiments.
3
Cell Culture and Virus Isolation Protocol
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IPI-2I cells (porcine intestinal epithelial cells), BHK-21 cells, and HeLa cells were obtained from the China Center for Type Culture Collection (Wuhan, China). LLC-PK1 cells were acquired from the American Type Culture Collection (ATCC CL-101; Manassas, VA, USA). IPI-2I, BHK-21, and LLC-PK1 cells were cultured in Dulbecco’s modified Eagle medium with 10% fetal bovine serum (Invitrogen, USA). HeLa cells were grown in RPMI 1640 medium with 10% fetal bovine serum. These cells were maintained in 5% CO2 at 37 °C. PDCoV strain CHN-HN-2014 (GenBank accession no. KT336560)10 (link) and TGEV strain WH1 (GenBank accession no. HQ462571) were isolated in 2014 and 2010, respectively, in China. Mouse monoclonal antibodies against TGEV M, TGEV N or PDCoV S, PDCoV N were created in-house. APN antibody was purchased from ABclonal (China). An anti-Flag rabbit polyclonal antibody (MBL, Japan), Alexa Fluor 594-conjugated donkey anti-rabbit IgG (Santa Cruz, USA), and Alexa Fluor 488-conjugated donkey anti-mouse IgG (Santa Cruz, USA) were used for indirect IFA. Horseradish peroxidase-conjugated goat anti-mouse antibody (Beyotime, China) and horseradish peroxidase-conjugated goat anti-rabbit antibody (Beyotime, China) were applied in western blots.
4
Quantifying Tsp1a Protein Abundance
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Ovaries from the WT and homozygous tsp1a mutants (n = 5/genotype) at 180 dah were dissected and pooled. Total proteins were extracted and diluted to a final concentration of 20 mg/ml. Western blot was carried out as described previously [41 (link)]. Antibody against Tsp1a was diluted at 1:500. The abundance of α-tubulin was examined as a loading control using rabbit anti-α-tubulin (Proteintech, Wuhan, China) at 1:1000. Horseradish peroxidase-conjugated goat anti-rabbit antibody (Beyotime, Shanghai, China) was used as the secondary antibody at 1:1000. Immunofluorescence signal was detected with BeyoECLPlus Kit (Beyotime, Shanghai, China) and was visualized on a Fusion FX7 (Vilber Lourmat, East Sussex, France).
5
Investigating Cisplatin-Induced Apoptosis
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IQC (HY-N0768) and U0126 (HY-12031) were purchased from MedChemExpress (MJ, NJ, United States). Cisplatin was procured from Sigma Aldrich (St. Louis, MO, United States). Periodic Acid-Schiff (PAS) stain kit was ordered from Servicebio (G1008, Wuhan, China). Antibodies against Bax, Bcl-2, Caspase-3, β-actin, GAPDH and Horseradish peroxidase-conjugated Goat Anti-Mouse antibody were obtained from Proteintech Group (Rosemont, IL, United States). MDA assay Kit (S0131), GSH and GSSG Assay Kit (S0053), Reactive Oxygen Species Assay Kit (S0033S) and Horseradish peroxidase-conjugated Goat Anti-Rabbit antibody were provided by Beyotime Biotech (Nantong, China). p44/42 MAPK (ERK1/2) (4695S), Phospho-p44/42 MAPK (p-ERK1/2) (8544S), NF-κB p65 (p65) (8242S) and Phospho-NF-κB p65 (P-p65) (4025S) were obtained from Cell Signaling Technology (Beverly, MA, United States). NGAL antibody is provided by Abcam (Cambridge, MA, United States). Dimethyl sulfoxide (DMSO) was provided from Sigma-Aldrich Chemicals (St. Louis, MO, United States). DAB (SA-HRP) TUNEL Cell Apoptosis Detection Kit (G1507) was from Servicebio (Wuhan, China). All other chemicals used in the study were commercially available and of analytical grade.
6
Western Blot Analysis of CNR-1 in Mouse PrL
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Mice were decapitated under deep anesthesia, and the PrL was rapidly separated and frozen at −80°C until use. The protein was abstracted using lysate. Protein concentration was estimated using a bicinchoninic acid (BCA) kit (Beyotime, China), and 30 μg samples were loaded onto polyacrylamide-SDS gels. Gels were electrophoresed and then transferred to PVDF membranes. Membranes were blocked with a blocking buffer containing 3% bovine serum albumin (Sigma-Aldrich, USA) for 60 min and probed with antibodies directed toward CNR-1 (1:1,000, Santa Cruz, USA) or GAPDH (1:2,000, ABclonal, China) overnight at 4°C. Primary antibody staining was detected with horseradish peroxidase-conjugated goat anti-rabbit antibodies (1:10,000, Beyotime, China). Each group was immune-blotted in three independent experiments, and average optical density relative to the internal standard (GAPDH) was reported and analyzed (Bio-Rad, USA).
7
LMHFV Stimulation of MC3T3-E1 Cells
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MC3T3-E1 cells (2 ϫ 10 5 cells/well) subjected to 5 consecutive days of LMHFV stimulation were lysed in RIPA lysis buffer containing 0.1 mM phenylmethylsulfonyl fluoride protease inhibitor. Total cellular protein was quantified using the BCA assay (Beyotime Institute of Biotechnology, Shanghai, China) and then subjected to electrophoresis using 12% SDS-PAGE. The separated protein bands were transferred onto polyvinylidene difluoride (PVDF) membranes, which were probed with COL-1 (ab96723; 1:1,000), OCN (ab93876; 1:1,000), OPN (ab8448; 1:1,000), and histone H3 (ab1791; 1:2,000) polyclonal rabbit primary antibodies (Abcam, Cambridge, UK) at 4°C overnight, followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit antibodies (A0208; Beyotime Institute of Biotechnology, Shanghai, China) for 1 h at 37°C. The blots were detected using the enhanced chemiluminescence reagent (NCM Biotech, Suzhou, China), and the bands were quantified using the ImageJ software.
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