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Penta·His HRP Conjugate Kit
Cat. No. / ID: 34460
125 μl Penta·His HRP Conjugate, 5 g Blocking Reagent, 50 ml Blocking Reagent Buffer (10x concentrate)
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Kit
Penta·His HRP
RGS·His HRP
Anti·His HRP Conjugate Kits 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
✓ Fast and reliable (re)-ordering
Features
- High-sensitivity and high-speed detection of His-tagged proteins, also with RGS·His epitopes
- No need for secondary antibody
- Optimized blocking reagent supplied
Product Details
Anti·His HRP Conjugate Kits, including the RGS·His HRP Conjugate Kit (His tag conjugates) and Penta·His HRP Conjugate Kit (6xHis tag conjugates), are intended for sensitive detection of recombinant proteins carrying His tags, without the need for secondary antibodies. They consist of anti·His antibodies conjugated to horseradish peroxidase (HRP). In detection procedures, the anti·His antibody binds to a protein's His tag and the complex is localized using chemiluminescent or chromogenic substrates that react with HRP.
Performance
| Feature | RGS·His HRP Conjugate Kit | Penta·His HRP Conjugate Kit |
|---|---|---|
| Detection | N-terminal, C-terminal and internal His tags (containing the epitope RGSHHHH) | N-terminal, C-terminal and internal 6xHis tags |
| Sensitivity in dot blots* | 10 pg | 10 pg |
| Sensitivity in western blots* | 50 pg | 50 pg |
| Antibody 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 | Stabilized stock solution | Stabilized stock solution |
| Working solution | 125–250 ml | 125–250 ml |
Principle
Anti·His HRP Conjugate Kits consist of the QIAexpress Anti·His Antibodies chemically coupled to the reporter enzyme horseradish peroxidase (HRP). They can be used for highly specific and sensitive, direct detection of any recombinant protein that has an accessible His tag. They eliminate the need for secondary antibodies, saving time in immunodetection procedures. Anti·His HRP Conjugate Kits can be used with chemiluminescent or chromogenic detection substrates.
Procedure
TheQIAexpress, Detection and Assay Handbook, supplied with all QIAexpress detection and assay products, contains detailed example protocols and guidelines for designing and performing assays.
Applications
Anti·His HRP Conjugate Kits, including the RGS·His HRP Conjugate Kit (His tag conjugate) and Penta·His HRP Conjugate Kit (6x His tag conjugate), recognize the same epitopes as their corresponding antibody and can be used for direct chemiluminescent or chromogenic detection of His-tagged proteins in:
- Western-, dot- and colony-blotting procedures
- ELISA and other immunoassay procedures
Comparison of the RGS·His HRP Conjugate Kit and Penta·His HRP Conjugate Kit
| Features | RGS·His HRP Conjugate Kit | Penta·His HRP Conjugate Kit |
|---|---|---|
| Conjugated enzyme | HRP | HRP |
| Detection | Secondary antibody required | Secondary antibody required |
| Epitope detected | RGSHHHH | HHHHH |
| Substrate for blot detection | Biotech ECL | 4C1N |
| Substrates for assay procedure | OPD | OPD |
| Tag | His tag | His tag |
Anti·His HRP Conjugate Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Resources
Safety Data Sheets (1)
Quick-Start Protocols (1)
Kit Handbooks (1)
Supplementary Protocols (1)
Certificates of Analysis (1)
FAQ
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
How can I avoid poor immunolocalization morphology with Anti-His Antibodies?
Can the Blocking Reagent supplied with the Penta-His HRP Conjugate be used with other antibodies?