For sensitive detection of His-tagged proteins

  • Detection of C-terminal, N-terminal, and internal His tags
  • Highly sensitive and specific detection of HHHHH epitopes

Monoclonal mouse Anti·His Antibodies are used to detect recombinant proteins carrying His tags. Each of the three variants detects a different His tag epitope.  Like all QIAGEN mouse monoclonal antibodies, they are prepared under serum-free conditions, which guarantees the absence of viruses, mycoplasma, and contaminating immunoglobulins.

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Penta·His Antibody, BSA-free (100 μg)
100 μg mouse anti-(H)5 (lyophilized, BSA-free, for 1000 ml working solution)
The Penta·His Antibody, BSA-free is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Sensitivity of QIAexpress Anti·His Antibodies.|His-tagged thioredoxin detected by Penta·His Antibody in yeast cells.|Identification of positive expression clones by colony blotting.|Immunohistochemistry with Penta·His Antibodies.|
Detection of 6xHis-tagged proteins (A–H, see table "Proteins tested with QIAexpress Anti·His Antibodies") with QIAexpress Anti·His Antibodies. Indicated amounts of pure 6xHis-tagged protein were applied to a nitrocellulose membrane, and detection was carried out with the Anti·His primary antibody indicated diluted 1/2000, followed by chromogenic detection with AP-conjugated rabbit anti-mouse IgG and NBT/BCIP.

 |The indicated amounts of purified 6xHis-tagged thioredoxin mixed with crude extract from yeast cells (5 µg total protein per lane) were applied to a 15% SDS-polyacrylamide gel. After electrophoresis and western transfer, 6xHis-tagged proteins were detected with Penta·His Antibody followed by chemiluminescent detection with HRP-conjugated rabbit anti-mouse IgG and ECL detection reagent. 6xHPL: 6xHis Protein Ladder.

 |Colony blot detection of positive expression clones with Penta·His Antibody. A BamHI–SalI fragment encoding a DNA polymerase was cloned into pQE-31, transformed into E. coli M15[pREP4], grown overnight at 37°C, replica-plated onto a nitrocellulose membrane, and treated according to the protocol in The QIAexpressionist with NBT/BCIP detection. Colonies of positive expression clones appear much darker than negative clones.

 |Immunolocalization of 6xHis-tagged β-galactosidase in murine NIH/3T3 fibroblast cells. Following transfection, tagged protein was detected using Penta·His Antibody followed by FITC/conjugated goat anti-mouse IgG antibody and visualized by fluorescence microscopy.


Performance of Penta·His Antibody BSA-free
Detection: N-terminal, C-terminal, and internal 6xHis tags
Sensitivity in dot blots:* 0.5 ng
Isotype: Mouse IgG1
Cross-reactivity: Negligible with crude E. coli, yeast, mammalian, or insect cell lysates
Form: Lyophilized
Working solution: 1000 ml
* Detection using chromogenic substrate. Sensitivity may vary with different proteins.

Many different proteins have been detected, via their His tags, using the QIAexpress Anti·His Antibodies (see table). For the sensitivity of the three Anti·His Antibodies in detecting this panel of proteins see the figure Sensitivity of QIAexpress Anti·His Antibodies)


Proteins tested with QIAexpress Anti·His Antibodies

Protein Tag locationSequence
C Thioredoxin N-terminus MRGSHHHHHHGSSDKIIHLT...
F Chaperonin C-terminus ...AKIAKDGSRSHHHHHH
G DNA Polymerase C-terminus ...EDWLSAKERSHHHHHH

For other experiments with  Anti·His Antibodies see figures Identification of positive expression clones by colony blotting and His-tagged thioredoxin detected by Penta·His Antibodies in yeast cells

All QIAexpress mouse monoclonal antibodies are produced under conditions that guarantee the highest purity and activity. Hybridoma cell culture is carried out under serum-free conditions, ensuring that the preparations are free of viruses, mycoplasma, and contaminating immunoglobulins. Purification by adsorption chromatography is performed entirely at physiological pH, yielding preparations with the highest proportion of native antibody. See figure Immunohistochemistry with Penta·His Antibodies.

QIAexpress Detection Systems allow efficient detection of His-tagged proteins and are ideal for many applications, including:

Colony, dot, and western blotting procedures 
Screening for positive expression clones 
Monitoring expression levels and stability of His-tagged proteins 
Immunoprecipitation and ELISA
Detection Secondary antibody required
Epitope detected HHHHH
Sensitivity in dot blots (chemiluminescent detection) 10 pg
Sensitivity in dot blots (chromogenic detection) 0.5 ng
Sensitivity in Western blots (chemiluminescent detection) 50 pg
Sensitivity in Western blots (chromogenic detection) 2 ng
Substrate for blot detection Dependent on secondary antibody
Substrates for assay procedure Dependent on secondary antibody
Tag His-tag

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Kit Handbooks
For Anti·His Antibodies, Anti·His HRP Conjugates, Penta·His™ Alexa Fluor® Conjugates, Penta·His Biotin Conjugate, Ni-NTA Conjugates, Tag·100™ Antibody, Streptavidin–R-PE, 6xHis Protein Ladder, Ni-NTA HisSorb™ Strips and Plates 
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