For sensitive detection of His-tagged proteins with HHHH epitopes

  • Detection of C-terminal, N-terminal, and internal His tags
  • Highly specific and sensitive detection
The Tetra·His Antibody is supplied as 100 µg of mouse monoclonal antibody (IgG1), as BSA-free, lyophilized powder. It can detect C-terminal, N-termal, and internal His tags. The Tetra·His Antibody has negligible cross-reactivity with crude E. coli, yeast, insect, or mammalian cell lysates.
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Product Cat. no. List price:
Tetra·His Antibody, BSA-free (100 µg)
100 µg mouse anti-(H)4 (lyophilized, BSA-free, for 1000 ml working solution)
34670
$1,161.00
The Tetra·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 anti·His antibodies.|
Detection of 6xHis-tagged proteins with Anti·His Antibodies (Tetra·His Antibody in the center). A: DHFR; B: DHFR; C: thioredoxin; D: TNF-α; E: TFIIAγ; F: chaperonin; G DNA polymerase; H: TFIIAαß; for tag location and sequences detected see table "Proteins detected 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.
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Performance

The sensitivity of Tetra·His Antibody in western blots is up to 2 ng, and up to 0.5 ng in dot blots (see figure "Sensitivity of Anti·His Antibodies"), when carrying out detection using a chromogenic substrate (sensitivity may vary with different proteins).

Proteins detected with QIAexpress Anti∙His Antibodies.
    Protein  Tag location Sequence
A     DHFR  N-terminus                                MRGSHHHHHHGSVRPLNCIV..........
B     DHFR  C-terminus            .........VYEKKDGSRSHHHHHH
C     Thioredoxin  N-terminus                                MRGSHHHHHHGSSDKIIHLT..........
D     TNF-α  N-terminus                                    MRHHHHHHQSTESMIRDV.........
E     TFIIAγ  N-terminus                                MGSSHHHHHHSSGLVPRGSH.....
F     Chaperonin  C-terminus            ..........AKIAKDGSRSHHHHHH
G

    DNA polymerase

 C-terminus            ..........EDWLSAKERSHHHHHH
H

    TFIIAαß

 Internal            ......QQQHQPQQQQHHHHHHHQQAQPQQTVP....

Principle

The QIAexpress System provides materials for the expression, purification, detection, and assay of 6xHis-tagged proteins. For detection, 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.

Tetra·His Antibody allows efficient detection of His-tagged proteins, and is ideal for many applications, including colony, dot, and western blotting procedures (see figure"Sensitivity of Anti·His Antibodies"), screening for positive expression clones, and monitoring expression levels and stability of 6xHis-tagged proteins. Tetra·His Antibody combined with conjugates has many additional useful applications, including immunoprecipitation, ELISA, immunocytochemistry, immunohistochemistry, protein localization and targeting studies, and LabMAP bead-based assays.

Applications

Tetra·His Antibody allows efficient detection of His-tagged proteins and is 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
  • Immunocytochemistry and immunohistochemistry
Feature
Specifications
Detection Secondary antibody required
Epitope detected HHHH
Sensitivity in dot blots -chemiluminescent substrate- 10 pg
Sensitivity in dot blots -chromogenic substrate- 0.5 ng
Sensitivity in western blots - Chemiluminescent substrate 50 pg
Sensitivity in western blots -chromogenic substrate- 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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Handbooks & Protocols
1
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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Kit Handbooks
1
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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References
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