Recombinant Protein Detection

KPL HisDetector™ Nickel Conjugates and Western Blot Kits eliminate the time and expense of using anti-His antibodies. HisDetector products utilize nickel-NTA conjugates which bind with high affinity to poly-histidine residues. They enable simple, direct detection of His-tagged proteins without the need for primary and secondary antibodies.

Recombinant protein detection at its best

  • The high affinity of histidine for nickel (Ni2+) ions, allows nickel-NTA (nitrilotriacetic acid) conjugates to bind directly to the His-tag (see figure on right)  
  • Detection is achieved with either chemiluminescent or colorimetric substrates
  • Nickel ions have six electron coordination bonds. NTA binds to four of these bonds (blue)
  • Two ligand binding sites (red) are then available to readily coordinate with His-tagged proteins (see figure on right)

Direct detection of His-tagged proteins with nickel conjugates greatly speeds assay times. Western Blot protocols take only 2.5 hours, saving several hours over traditional antibody detection methods. Assay optimization is minimal when compared to antibody methods that require optimization of multiple antibodies.

Speed, ease-of-use, proven performance

Picogram level sensitivity in Western blotting assays with very low background allows for specific detection of his-tagged proteins using either Nickel-HRP or Nickel-AP conjugates (Figure 2Figure 3). His-tagged protein ladder (lanes a-e) and E. coli crude lysate containing His-tagged B-gal (lanes 1-3) were detected in Western Blot using Nickel-HRP and Nickel-AP and corresponding chemiluminescent and colorimetric substrates.

Figure 2 - Nickel-HRP                          Figure 3 - Nickel-AP
Nickel-HRP     Nickel-AP

Product List

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Material # Reference # Product Name Size List Price Unit



HisDetector™ Western Blot Kit, HRP Colorimetric 40 blots Contact Us EA



HisDetector™ Western Blot Kit, HRP Chemiluminescent 40 blots Contact Us EA



HisDetector™ Western Blot Kit, AP Colorimetric 40 blots Contact Us EA



HisDetector™ Nickel-HRP 0.1 mL Contact Us EA



HisDetector™ Nickel-AP 1 mL Contact Us EA