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.
100 μg mouse anti-(H)5 (lyophilized, BSA-free, for 1000 ml working solution)
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 of Penta·His Antibody BSA-free
N-terminal, C-terminal, and internal 6xHis tags
Sensitivity in dot blots:*
Negligible with crude E. coli, yeast, mammalian, or insect cell lysates
* 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
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
Secondary antibody required
Sensitivity in dot blots (chemiluminescent detection)
Sensitivity in dot blots (chromogenic detection)
Sensitivity in Western blots (chemiluminescent detection)
Sensitivity in Western blots (chromogenic detection)