Qiagen, 34660, Penta·His Antibody, BSA-free (100 µg)

100 µg mouse anti-(H)5 (lyophilized, BSA-free, for 1000 mL working solution)

Features

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

Product Details

Monoclonal mouse Anti·His Antibodies (5xHis antibodies and 4xHis antibodies), like the RGS·His Antibody, BSA-free, Tetra·His Antibody, BSA-free and Penta·His Antibody, BSA-free are used to detect recombinant protein 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.

Performance

Feature: RGS·His Antibody, BSA-free
Detection: N-terminal His tags (containing the epitope RGSHHHH)
Epitope detected: RGSHHHH
Sensitivity in dot blots*: 0.5 ng
Sensitivity in western blots*: 2 ng
Isotype: Mouse IgG1
Cross-reactivity: Negligible with crude E. coli , yeast, mammalian or insect cell lysates
Form: Lyophilized
Working solution: 1000 mL

Protein: Tag location
A: DHFR
B: DHFR
C: Thioredoxin
D: TNF-α
E: TFIIAγ
F: Chaperonin
G: DNA polymerase
H: TFIIAαß

Many different proteins have been detected, via their His tags, using QIA express Anti·His Antibodies (5xHis antibodies and 4xHis antibodies) (see table “Performance of Anti∙His Antibodies”). For the sensitivity of the three Anti·His Antibodies in detecting this panel of proteins see the figure “Sensitivity of QIA express Anti·His Antibodies” ). Anti·His Antibodies enable identification of positive expression clones by colony blotting (see figure “Identification of positive expression clones by colony blotting” ) and detection of His-tagged thioredoxin (see figure “His-tagged thioredoxin detected by Penta·His Antibody in yeast cells” ). The antibodies are also ideally suited for use in immunohistochemistry procedures (see figure “Immunohistochemistry with Penta·His Antibodies” ).

Performance of Anti∙His Antibodies

Proteins detected with QIAexpress Anti∙His Antibodies

Principle

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

Procedure

The QIA express , Detection and Assay Handbook , supplied with all QIA express detection and assay products, contains detailed example protocols and guidelines for designing and performing assays.

Applications

- 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

QIA express Detection Systems, including RGS·His Antibody, BSA-free, Tetra·His Antibody, BSA-free and Penta·His Antibody, BSA-free allow efficient detection of His-tagged proteins and are ideal for many applications, including: