Alveolar macrophages/HUVECs were lysed in the RIPA buffer (Thermo Scientific, Rockford, USA) supplemented with 1% PMSF and 1% protein phosphatase inhibitor mixture (P1260, Applygen, Beijing, China). The concentration of the proteins was measured with the Bicinchoninic Acid Kit (Thermo Scientific, Rockford, USA). Rabbit anti-TLR2, TLR4, and phosphorylated p65 unit of NF-κB antibodies were obtained from Cell Signaling Technology. Horseradish peroxidase- (HRP-) conjugated goat anti-rabbit antibodies were purchased from Bio-Rad. ECL-Plus Chemiluminescent Reagent (Thermo Scientific, Rockford, USA) was used to visualize the specific proteins. Relative concentration of proteins was quantitated using the Image J Software (National Institutes of Health, Bethesda, USA).
Horseradish peroxidase hrp conjugated goat anti rabbit antibodies
Manufactured by Bio-Rad
Horseradish peroxidase- (HRP-) conjugated goat anti-rabbit antibodies are secondary antibodies that bind to rabbit primary antibodies. These antibodies are conjugated to the enzyme horseradish peroxidase, which can be used for detection in various immunoassays and immunohistochemical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Bicinchoninic acid kit, by Thermo Fisher Scientific (1 mentions)
Ecl plus chemiluminescent reagent, by Thermo Fisher Scientific (1 mentions)
Immpact novared peroxidase hrp substrate, by Vector Laboratories (1 mentions)
Eukitt, by Merck Group (1 mentions)
Aperio imagescope, by Leica (1 mentions)
2 protocols using horseradish peroxidase hrp conjugated goat anti rabbit antibodies
1
TLR2, TLR4, and NF-κB Protein Quantification
2
Phosphorylated HSL Immunohistochemistry in Rat Liver
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Immunohistochemistry for phosphorylated HSL (anti-pHSL; Supplementary Table S2) was performed on paraffin-embedded rat liver sections as previously described [29] (link). Briefly, after deparaffinization antigen retrieval was performed by microwave irradiation in citrate buffer (10 mmol/L), pH 6.0 and blocking of endogenous peroxidase with 0.3% H 2 O 2 for 30 min. After blocking (1% BSA for 30 min), tissue was incubated with polyclonal-rabbit-phospho-HSL (Ser660) primary antibody (1:50 dilution in 1% BSA) overnight at 4 °C in a humidity chamber. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibodies (1:50; #170-6515, Bio-Rad) was used as secondary antibody. Slides were stained with ImmPact NovaRED Peroxidase (HRP) substrate (cat# SK-4805, Vector Laboratories, The Netherlands) for 11 min, and hematoxylin was used as a counter nuclear stain (2 min at RT). Finally, slides were dehydrated and mounted with Eukitt®, (Sigma-Aldrich). Slides were scanned using a nanozoomer 2.0 HT digital slide scanner (C9600-12, Hamamatsu Photonics, Hamamatsu, Japan) and analyzed with Aperio ImageScope (version 11.1, Leica Microsystems, Amsterdam, The Netherlands).
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