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Penta·His Antibody, BSA-free

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.

Cat No./ID: 34660
Penta·His Antibody, BSA-free (100 μg)
€618.00
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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.

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.


 

Performance
Performance of Penta·His Antibody BSA-free
PropertyValue
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
A DHFR N-terminus MRGSHHHHHHGSVRPLNCIV...
B DHFR C-terminus ...VYEKKDGSRSHHHHHH
C Thioredoxin N-terminus MRGSHHHHHHGSSDKIIHLT...
D TNFα N-terminus MRHHHHHHQSTESMIRDV...
E TFIIAγ N-terminus MGSSHHHHHHSSGLVPRGSH...
F Chaperonin C-terminus ...AKIAKDGSRSHHHHHH
G DNA Polymerase C-terminus ...EDWLSAKERSHHHHHH
H TFIIAαβ Internal ...QQQHQPQQQQHHHHHHQQAQPQQTVP...

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

Principle
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.
Applications

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
Features
Specifications
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 (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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