Anti·His HRP Conjugate Kits

For sensitive, direct detection of His-tagged proteins

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
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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)
$564.00
Kit
Penta·His HRP
RGS·His HRP
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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

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

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

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

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 
Supplementary Protocols (1)

FAQ

What epitopes do the Anti-His Conjugates and Anti-His Antibodies recognize?

The table below lists the epitopes recognized by the Anti-His Conjugates and Anti-His Antibodies. 'H' designates a histidine residue, while 'X' designates any amino acid other than histidine. Note that individual 6xHis-tagged proteins are often recognized better by one Anti-His Antibody than by the others, possibly because of subtle differences in the exact conformation of the 6xHis tag and other parts of the protein in the vicinity of the tag. To choose the optimal antibody for your specific 6xHis-tagged protein and application, QIAGEN offers the Anti-His Antibody Selector Kit. The kit includes 3 µg of each of the three antibodies for direct comparison and economical selection of the one that gives the best results.

Epitopes recognized by QIAexpress Anti-His HRP Conjugates and Anti-His Antibodies


Peptide Penta-His Tetra-His RGS-His
XHHHHHHX
+ +
XXHHHHHX
+ +
XHXHHHHX
+
XHHHHXHX
+
XRGSHHHHX
+ +
XRGSHHHHHHX
+ + +
XHXHHXHX

Note that the presence of specific residues N- or C-terminal to the epitope is not required.

FAQ ID -170
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?
Insufficient fixation of samples leads to poor morphological preservation. If using methanol or acetone try fixation with paraformaldehyde. If 2% paraformaldehyde is insufficient, increase fixative concentration to 4% and increase incubation time. Keep in mind that too harsh fixation can lead to functional alteration of the 6xHis epitope and loss of recognition by the Anti·His Antibodies.immunofluorescence
FAQ ID -200
Can the Blocking Reagent supplied with the Penta-His HRP Conjugate be used with other antibodies?
Yes, the Blocking Reagent supplied with our Penta-His HRP Conjugate can also be used with other antibodies.
FAQ ID -1073