An innovative probe and kit for His-tagged protein detection.
Thermo Scientific HisProbe-HRP is a nickel (Ni2+)-activated derivative of horseradish peroxidase (HRP) that enables direct, IMAC-based detection of His-tagged proteins and other histidine-rich proteins in Western blots and microplates.
Unlike anti-6xHis antibodies, HisProbe-HRP can recognize polyhistidine tags independent of adjacent tags. The active ligand in HisProbe-HRP is a tridentate chelator that allows Ni2+ to be bound in active form for subsequent interaction and detection fo target molecules. The active chelator has binding capabilities similar to iminodiacetic acid (IDA) when used for immobilized metal affinity chromatography (IMAC). The Thermo Scientific SuperSignal West HisProbe Kit includes blocking buffers and chemiluminescent substrate for sensitive Western blot detection via the HisProbe-HRP Conjugate.
Highlights:
Fast – His-tag detection is achieved with a one-step probe incubation that eliminates the need to run a lengthy two-step primary/secondary antibody reaction protocol
Economical – no need to purchase expensive primary/secondary antibodies; considerably less costly than anti-6xHis-tag antibodies
High specificity – HisProbe-HRP can recognize his-tags independent of adjacent tags.
Less steric hindrance – HisProbe-HRP is smaller than antibodies and detects his tagged fusion proteins that are often undetectable when using some anti-his-tag antibodies
Product Details:
Mechanism for detection with Thermo Scientific HisProbe-HRP probe.
Comparison of His-tagged fusion protein detection methods. Panel A. The HisProbe-HRP provided high specific binding for all three His-tagged proteins and low background. Panel B. Anti-6xHis antibody failed to recognize two of the three His-tagged proteins and caused considerable background.
References:
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Wagner C. et al. Dimerization of NO-sensitive guanylyl cyclase requires the alpha1 N terminus. J. Biol. Chem., Mar 2005
Sundberg-Smith L., et al. Adhesion Stimulates Direct PAK1/ERK2 Association and Leads to ERK-dependent PAK1 Thr 212 Phosphorylation. J. Biol. Chem., Jan 2005; 280: 2055 – 2064.
