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Horseradish peroxidase hrp conjugated second antibody

Manufactured by Merck Group
Sourced in Japan

Horseradish peroxidase (HRP)-conjugated second antibody is a laboratory product used as a detection reagent. It contains an enzyme, horseradish peroxidase, which is chemically conjugated to a secondary antibody. This product is used in various immunoassay techniques to amplify and detect target analytes.

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2 protocols using horseradish peroxidase hrp conjugated second antibody

1

Comprehensive Chemical Reagents Inventory

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Generally, all chemicals were obtained from FUJIFILM-Wako (Tokyo, Japan), unless stated otherwise. Fetal bovine serum (FBS) for cell culture was purchased from Gibco (GIBCO BRL, Grand Island, NY, USA). Cell Counting Kit-8 (CCK-8) and Cytotoxicity lactate dehydrogenase (LDH) Kit-WST were purchased from Dojindo Molecular Technologies (Rockville, MD, USA). Cellular ROS Kit (#ab113851) was obtained from Abcam (Tokyo, Japan). Nuclear Extraction (NE)-Kit and human TNF-alpha ELISA Ready-SET Go Kit (#88-7324) were obtained from eBiosciences (San Diego, CA, USA), respectively. Pierce bicinchoninic acid-(BCA) protein Kit and Enhanced chemiluminescent-(ECL) Western blotting substrate were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Bovine serum albumin-(BSA) fraction V was purchased from Roche (Mannheim, Germany). All antibodies for Western blot analysis were purchased from Cell Signaling Technology (Tokyo, Japan), except anti-Nrf2 (Proteintech Corp., Tokyo, Japan), anti-p65 NF-ĸB, and horseradish peroxidase (HRP)-conjugated second antibody (Sigma-Aldrich, Burlington, MA, USA), anti-rabbit (IgG) Alexa FluorTM 488-conjugate (Alexa-488), anti-rabbit (IgG) Alexa FluorTM 594-conjugate (Alexa-594) (Molecular Probes, Invitrogen, CA, USA). DAPI (4′,6-diamino-2-phenylindole) dye used for the nuclei staining was purchased from Molecular Probes (Eugene, OR, USA).
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2

Western Blot Analysis of Viperin and ZIKV NS5

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Cells were solubilized in lysis buffer containing 2% SDS, 25 mM Tris, 150 mM NaCI, 1% NP40. Cell lysates were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with anti-HA mAb (Covance) to detect viperin in CHME3.viperin cells, anti-viperin mAb (Milipore) to detect viperin in R848-treated human primary cells, rabbit anti-ZIKV NS5 antiserum, and anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase, Sigma) MAb. The membranes were washed and then probed with goat anti-mouse or anti-rabbit horseradish peroxidase (HRP)-conjugated second antibody (Sigma) and visualized using HRP substrate (Pierce) on an Odyssey Fc dual-mode imaging system (Li-Cor).
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