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HisProbe-HRP and SuperSignal West HisProbe Kit


An innovative probe and kit for His-tagged protein detection.

SuperSignal West HisProbe Kit

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:

HisProbe-HRP mechanism of action
Mechanism for detection with Thermo Scientific HisProbe-HRP probe.

Using HisProbe-HRP shows high specific binding for all three His-tagged proteins and low background. Anti-6xhis antibody failed to recognize two of the three His-tagged proteins and caused considerable background.
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:

  1. Segawa H., et al. Reconstitution of GDP-mannose Transport Activity with Purified Leishmania LPG2 Protein in Liposomes, J. Biol. Chem., Jan 2005; 280: 2028 – 2035.
  2. Adler J. and Bibi E. Determinants of Substrate Recognition by the Escherichia coli Multidrug Transporter MdfA Identified on Both Sides of the Membrane, J. Biol. Chem., Mar 2004; 279: 8957 – 8965
  3. Adler J. and Bibi E., Promiscuity in the Geometry of Electrostatic Interactions between the Escherichia coli Multidrug Resistance Transporter MdfA and Cationic Substrates. J. Biol. Chem., Jan 2005; 280: 2721 – 2729
  4. Wagner C. et al. Dimerization of NO-sensitive guanylyl cyclase requires the alpha1 N terminus. J. Biol. Chem., Mar 2005
  5. 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.

Related Resources:

His-tagged fusion protein production and purification technical guide

Related Products:

His-tagged Fusion Protein Purification Resins and Kits
Nickel Coated Plates
Copper Coated Plates
Anti-Tag Antibodies


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
15165 HisProbe-HRP Conjugate
Formulation: Lyophilized powder from protein in MES buffer, pH 5.5
Sufficient For: Preparing approx. 2L of 1X probing solution
2mg Product Instructions for product #15165 HisProbe-HRP Conjugate MSDS for product #15165 HisProbe-HRP Conjugate Certificate of Analysis for product #15165 HisProbe-HRP Conjugate $186.00

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15168 SuperSignal West HisProbe Kit
Formulation: Multi-component kit
Sufficient For: 62 mini blots or 5000cm2 membrane

Kit Contents:
HisProbe-HRP Conjugate, 2mg
SuperSignal West Pico Luminol/Enhancer Solution, 250mL
SuperSignal West Pico Stable Peroxide Solution, 250mL
Blocker BSA in TBS (10X), 125mL
TBS Packs (each makes 500mL), 10 packs
Surfact-Amps 20 Detergent Solution, 6 x 10mL
685mL kit Product Instructions for product #15168 SuperSignal West HisProbe Kit MSDS for product #15168 SuperSignal West HisProbe Kit Certificate of Analysis for product #15168 SuperSignal West HisProbe Kit $441.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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