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RGS·His Antibody, BSA-free (100 µg)
Cat. No. / ID: 34650
100 µg mouse anti·RGS(H)4 (lyophilized, BSA-free, for 1000 mL working solution)
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Tag
RGS·His
Tetra·His
Penta·His
Anti·His Antibodies, BSA-free are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
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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
Many different proteins have been detected, via their His tags, using QIAexpress 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 QIAexpress 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
| Feature | RGS·His Antibody, BSA-free | Tetra·His Antibody, BSA-free and Penta·His Antibody, BSA-free |
|---|---|---|
| Detection | N-terminal His tags (containing the epitope RGSHHHH) | N-terminal, C-terminal and internal 4xHis (Tetra·His Antibody, BSA-free) or 5xHis tags (Penta·His Antibody, BSA-free) |
| Epitope detected | RGSHHHH | HHHH (Tetra·His Antibody, BSA-free) or HHHHH (Penta·His Antibody, BSA-free) |
| Sensitivity in dot blots* | 0.5 ng | 0.5 ng |
| Sensitivity in western blots* | 2 ng | 2 ng |
| Isotype | Mouse IgG1 | Mouse IgG1 |
| Cross-reactivity | Negligible with crude E. coli, yeast, mammalian or insect cell lysates | Negligible with crude E. coli, yeast, mammalian or insect cell lysates |
| Form | Lyophilized | Lyophilized |
| Working solution | 1000 mL | 1000 mL |
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.... |
See figures
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.
Procedure
The QIAexpress, Detection and Assay Handbook, supplied with all QIAexpress detection and assay products, contains detailed example protocols and guidelines for designing and performing assays.
Applications
QIAexpress 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:
- 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
Supporting data and figures
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.
Resources
Safety Data Sheets (1)
Kit Handbooks (1)
Supplementary Protocols (1)
Certificates of Analysis (1)
Publications
Localization of the gD-binding region of the human herpes simplex virus receptor, HveA.
J Virol; 2001; 75 (1):171-80 2001 Jan PMID:11119586
Multiple sulfatase deficiency is caused by mutations in the gene encoding the human C(alpha)-formylglycine generating enzyme.
Cell; 2003; 113 (4):435-44 2003 May 16 PMID:12757705
Screening for soluble expression of recombinant proteins in a 96-well format.
Anal Biochem; 2001; 297 (1):79-85 2001 Oct 1 PMID:11567530
A human cDNA expression library in yeast enriched for open reading frames.
Genome Res; 2001; 11 (10):1730-5 2001 Oct PMID:11591650
Phosphorylation of the vesicle-tethering protein p115 by a casein kinase II-like enzyme is required for Golgi reassembly from isolated mitotic fragments.
J Cell Biol; 2000; 150 (3):475-88 2000 Aug 7 PMID:10931861
FAQ
How should I dissolve the lyophilized Anti-His and Tag Antibodies?
What epitopes do the Anti-His Conjugates and Anti-His Antibodies recognize?
Which Anti-His Antibody is the most sensitive for my protein of interest?
Penta-His and Tetra-His Antibody bind to proteins that have 6xHis tags regardless of the surrounding amino-acids, even when the tag is partially hidden. Subtle differences in the antigen recognition sites of these antibodies mean that some 6xHis-tagged proteins are detected with higher sensitivity by one Anti-His Antibody than by the others (see figure below). Therefore, to achieve optimum results, we recommend comparison of these antibodies in parallel for new applications and proteins. QIAGEN offers the Anti-His Antibody Selector Kit for this purpose.
Sensitivity of QIAexpress Anti·His Antibodies
Note: RGS His Antibody regonizes the RGS(His)4 epitope that is found on proteins endcoded by most pQE vectors as well as several pRSETan pBlue BacHis -vectors from Invitrogen.
FAQ ID -172
What are the dissociation constants for the Anti-His Antibodies?
Are QIAGEN monoclonal Anti His Antibodies glycosylated?
How is an antibody epitope determined?
What blocking reagents do you recommend for chemiluminescent detection using Penta-, Tetra-, and RGS-His antibodies?
What is the difference between BSA-free and BSA-containing RGS-His Antibody?
How should I store Anti·His and Tag·100 Antibodies?