OptiMEM reduced serum medium and Lipofectamine 2000 reagent were purchased from Gibco Life Technologies (Grand Island, NY, USA). Protease inhibitor cocktail tablets (Complete Mini, EDTA-free) were purchased from Roche (Mannheim, Germany). Primary antibodies utilized were polyclonal rabbit anti-human TRPM2 antibody (Cat. #A300-414A, Bethyl Laboratories, Montgomery, TX, USA), polyclonal rabbit anti-human β-actin (Cat. #600-401-886, Rockland Immunochemicals, Limerick, PA, USA), polyclonal rabbit anti-human manganese superoxide dismutase (MnSOD) (Cat. #06-984, Millipore, Billerica, MA, USA), and monoclonal mouse anti-human Lamin B2 clone LN43 (Cat. #MA1-06104, Thermo Fisher Pierce, Pittsburgh, PA, USA). The secondary antibodies, horseradish peroxidase (HRP)-conjugated goat anti-rabbit and HRP-conjugated rabbit anti-mouse were purchased from Sigma (St. Louis, MO, USA). 2-Aminoethoxydiphenyl borate (2-APB), maintained as a 75 mM stock solution in dimethyl sulfoxide (DMSO), and 30% hydrogen peroxide solution were purchased from Sigma. The Fluo-4 NW Calcium Assay kit was purchased from Life Technologies. Comet Assay kit, which includes alkaline lysis solution, LMAgarose, 2-well CometSlides, SYBR Green, and EDTA, was purchased from Trevigen (Gaithersburg, MD, USA). CytoScan WST-1 cell proliferation assay was purchased from VWR International (Radnor, PA, USA).
Horseradish peroxidase hrp conjugated goat anti rabbit
Manufactured by Merck Group
Sourced in United States
Horseradish peroxidase (HRP)-conjugated goat anti-rabbit is a secondary antibody used in various immunoassay techniques. It consists of goat-derived antibodies that specifically bind to rabbit primary antibodies, with horseradish peroxidase enzyme conjugated to facilitate signal detection.
Automatically generated - may contain errors
Lab products found in correlation
Fluo 4 nw calcium assay kit, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
2 aminoethoxydiphenyl borate 2 apb, by Merck Group (1 mentions)
Lmagarose, by Bio-Techne (1 mentions)
2 well cometslides, by Bio-Techne (1 mentions)
Sybr green, by Bio-Techne (1 mentions)
Opti mem reduced serum medium, by Thermo Fisher Scientific (1 mentions)
Comet assay kit, by Bio-Techne (1 mentions)
Rabbit anti pig igg, by Merck Group (1 mentions)
3 protocols using horseradish peroxidase hrp conjugated goat anti rabbit
1
Optimized Transfection and Cellular Assay
2
Western Blot Analysis of PDCoV S1 Protein
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Purified PDCoV S1 protein with an Fc-tag was subjected to 12% SDS-PAGE and then transferred to a nitrocellulose (NC) membrane. The NC membrane was blocked using 5% (w/v) nonfat dried milk in phosphate-buffered saline (PBS) at 37 °C for 1 h and then incubated with a rabbit monoclonal antibody (mAb) against the Fc tag (1:20,000) and a porcine polyclonal antibody (pAb) against PDCoV (1:500) at 37 °C for 1 h. After washing three times with PBS containing 0.05% Tween 20 (PBST), the membrane was incubated in PBS containing horseradish peroxidase (HRP)-conjugated goat anti-rabbit (1:10,000) (Sigma) or rabbit anti-pig IgG (1:10,000) (Sigma) at 37 °C for 1 h. Membranes were washed with PBST and incubated with substrate solution (PBST, 0.05% DAB and 0.006% H2O2).
3
Polyclonal Antibody Production against BmCPV VP7
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To prepare polyclonal antibody against VP7 proteins of BmCPV, the cDNA with primers listed in Table 1 was amplified with PCR. The amplified PCR products were ligated into the prokaryotic expression vector pET28(a). The recombinant prokaryotic expression vector pET28(a)-vp7 (Supplementary Fig. 1 A) was transformed into Escherichia coli strain BL21 cells and induced with 1 mmol/L isopropyl-β-d -thiogalactopyranoside (IPTG). After 4 h of induction, 1.5 ml of the transformed BL21 cells was collected for the recombinant protein detection. The proteins were detected with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting (Supplementary Fig. 1 B), using a primary antibody of rabbit anti-6 × His (Sigma) and a secondary antibody of horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Sigma). Recombinant protein (VP7) was purified from the 1 L BL21 cells with Ni-NTA (Ni2 +-nitrilotriacetate) resin (Invitrogen, Carlsbad, CA) using standard procedure. Mouse was immunized with recombinant protein (VP7) for preparing the VP7 polyclonal antibody. Booster injections were administered according to the manufacturer's instructions. The collected serum was stored at − 70 °C.
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