Protein from human and rodent skin was extracted in RIPA buffer containing phosphatases and proteases inhibitor cocktails (Roche). Protein concentrations were determined by the BCA protein assay kit (Pierce, Mississauga, ON, Canada). Proteins were resolved by SDS-PAGE followed by western blotting using the following antibodies at 1:1000 concentration: Caspase-1 (Cell Signaling, MA), cleaved Caspase-1 (Cell Signaling, MA), IL1β (Cell Signaling, MA), cleaved IL1β (Cell Signaling, MA), and GAPDH (Cell Signaling, MA). Species appropriate secondary antibodies conjugated to horseradish peroxidase (BioRad, Mississauga, ON, Canada) were used and proteins visualized by enhanced chemiluminescence using the BioRad ChemiDoc MP Imaging System. Band intensities were detected, normalized and quantified with the ChemiDoc and Image Lab 5.0 software (BioRad Laboratories, Hercules, CA). Antibody concentrations are expressed relative to GAPDH.
Antibodies conjugated to horseradish peroxidase
Manufactured by Bio-Rad
Sourced in Canada, United States
Antibodies conjugated to horseradish peroxidase are a type of laboratory reagent used for various analytical techniques. The horseradish peroxidase enzyme is covalently attached to the antibody, allowing it to be used as a detection system in applications such as Western blotting, ELISA, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Image lab 5, by Bio-Rad (1 mentions)
Proteases inhibitor cocktail, by Roche (1 mentions)
Chemidoc, by Bio-Rad (1 mentions)
Il 1β, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Cleaved caspase 1, by Cell Signaling Technology (1 mentions)
Cleaved il 1β, by Cell Signaling Technology (1 mentions)
Caspase 1, by Cell Signaling Technology (1 mentions)
Quantity one 4, by Bio-Rad (1 mentions)
Clarity western ecl, by Bio-Rad (1 mentions)
Tsg101, by Abcam (1 mentions)
2 protocols using antibodies conjugated to horseradish peroxidase
1
Quantitative Analysis of Skin Inflammasome
2
Extracellular Vesicle Protein Profiling
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15 μg protein isolated from the serum EV fraction was separated by SDS–PAGE (10% polyacrylamide) according to a standard protocol. The polypeptides were electroblotted onto nitrocellulose membrane (Immobilon®-FL PVDF membrane) and probed with a primary antibody overnight at 4° C. Alix (1:200; Santa Cruz sc-53538), Tsg101 (1:500; Abcam ab83), CD63 (1:5000; ABIN 1440014), CD79a (1:500; ABIN 2472431), CD20 (1:2000; ABIN 2717455), CD21 (1:500; ABIN 94032), CD45 (1 μg/ml; ABIN 6940460) antibodies were used as primary antibodies. Anti-goat (for CD63) or anti-mouse (for the rest) antibodies conjugated to horseradish peroxidase (Bio-Rad, Hercules, CA, USA) were used as secondary antibodies at a dilution of 1/5000 or 1/10000, respectively. Visualization was performed with a chemiluminescent reagent system (Clarity™Western ECL, Bio-Rad, Hercules, CA, USA). Blots were quantified by densitometry using Quantity One 4.6.2 software (Bio-Rad Ltd.).
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