Restriction enzymes were purchased from Promega (Madison, WI). Pfu DNA polymerase was supplied by Stratagene (Madison, WI). Ampicillin and isopropyl thio-β-D-galactopyranoside (IPTG) were obtained from TaKaRa Shuzo (Otsu, Shiga, Japan). The 7-amino-4-methylcoumarin (amc)-linked substrates L-D-amc, L-S-amc, L-V-amc, L-A-amc, L-F-amc, L-R-amc, and hippuryl-R were purchased from GL Biochem (Shanghai, China). The His-Tag™ monoclonal antibody and the rabbit anti-mouse peroxidase-conjugated secondary antibody were purchased from Merck (San Diego, CA). Other chemicals used were reagent-grade. The pET-15b vector and the E. coli strain were purchased from Novagen (Madison, WI).
His tag monoclonal antibody
Manufactured by Merck Group
Sourced in United States
His-Tag™ monoclonal antibody is a laboratory tool used to detect and purify proteins of interest. It is designed to bind to a specific amino acid sequence (the histidine tag) that can be genetically attached to the target protein, allowing for its identification and isolation from complex mixtures.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using his tag monoclonal antibody
1
Recombinant Protein Expression Protocol
2
Recombinant PkCSP Protein Characterization
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Crude protein lysate and purified recombinant PkCSP were resolved in 12% SDS-PAGE under reducing conditions using b-mercaptoethanol as the reducing agent. The gels were stained with coomassie brilliant blue to reveal the protein bands. Purified recombinant PkCSP was excised from SDS-PAGE gel and sent for protein identification using MALDI/TOF. Separated proteins were transferred onto polyvinylidene difluoride (PVDF) membrane and blocked overnight in 5% blocking buffer (1X tris-buffered saline (TBS) containing 5% skimmed milk) at 4°C. The membrane was probed with His-Tag® monoclonal antibody (EMD Millipore Corp., USA) diluted with 2.5% blocking buffer (1:2500 dilution) for one hour at room temperature. The membrane was washed three times with 0.2% TBS-T (1X TBS containing 0.2% Tween-20) and incubated with biotin-labelled goat anti-mouse IgG (1:2500 dilution) for one hour, followed by alkaline phosphataseconjugated streptavidin (1:2500 dilution) for one hour. Finally, nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (NBT/BCIP) was added in the dark to develop colour.
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