Miltenyi Biotec
Miltenyi Biotec home - back to start page
Miltenyi Biotec Miltenyi Biotec
Miltenyi Biotec
Miltenyi Biotec

Miltenyi Biotec
Gene expression profiling products
miRNA expression profiling products
a-Hyb™ Hybridization Station
KIR typing
navigationProtein isolation and analysis
Miltenyi 

Biotec
Isolation and analysis of epitope-tagged proteinsIsolation and analysis of epitope-tagged proteinsIsolation and analysis of epitope-tagged proteins
Miltenyi 

Biotec
Isolation of interacting molecules with a biotinylated probeIsolation of interacting molecules with a biotinylated probeIsolation of interacting molecules with a biotinylated probe
Miltenyi 

Biotec
Immunopurification with Protein A / Protein G MicroBeadsImmunopurification with Protein A / Protein G MicroBeadsImmunopurification with Protein A / Protein G MicroBeads
Miltenyi 

Biotec
Native transcription factor isolationNative transcription factor isolationNative transcription factor isolation
Miltenyi 

Biotec
Large scale recombinant protein isolationLarge scale recombinant protein isolationLarge scale recombinant protein isolation
Miltenyi 

Biotec
HIV/virus isolation
Mitochondria isolation
Transfected cell selection
MACS® Separators and Columns
Genomics services
Miltenyi Biotec
MACS eNewsletter
Subscribe to the
MACS eNewsletter
for regular product information via e-mail.
Miltenyi Biotec
Isolation and analysis of His-tagged proteins
Background
The polyhistidine (His) tag is an amino acid motif that consist of six histidine residues which are added at the C-terminus or N-terminus of recombinant protein for affinity purification.

High-quality monoclonal anti-His antibodies are offered by Miltenyi Biotec either coupled to µMACS™ MicroBeads for specific isolation of His-tagged proteins on an analytical scale or conjugated to biotin, FITC, or horseradish peroxidase (HRP) for analysis of recombinant His-tagged protein.
Isolation and detection of His-tagged proteins
Isolation of His-tagged proteins
The µMACS™ and MultiMACS™ His Isolation Kits take advantage of the well-established MACS® Technology; thus, allows fast and specific isolation of pure His-tagged proteins. The advantages of MACS Technology for protein isolation are:

Specificity – high-quality monoclonal antibodies coupled to MACS MicroBeads assure specific isolation of His-tagged proteins with low background.
Sensitivity – fast reaction kinetics of tiny (50 nm), non-sedimenting MACS MicroBeads allow sensitive isolation even when working with rare proteins.
Speed – less than 2 hours to high-purity His-tagged proteins!

The µMACS His Isolation Kit was developed for manual, low-throughput applications with the µMACS Separator.
The procedure can easily be upscaled with the MultiMACS His Isolation Kits to a semi- or fully-automated, high-throughput processing of up to 96 samples in parallel by utilizing the MultiMACS 96 Separator.

For medium- to large-scale purification of His-tagged proteins the ProCatch His Resin is recommended (fig. 1).


Detection of His-tagged proteins
The monoclonal Anti-His antibody is available with various conjugates. The fluorochrome-conjugated (FITC or PE) Anti-His antibodies enable an immunofluorescence analysis for example, by fluorescence microscopy or FACS (fig. 2). Various detection methods are applicable using the biotinylated Anti-His antibody in combination with appropriate streptavidin conjugates.

For specific and sensitive detection of His-tagged proteins by Western blot or ELISA analysis the Anti-His-HRP and the Anti-His-HRP (C-term.) are available. Directly coupled to horseradish peroxidase (HRP), both antibodies simplifiy these immunological assays as incubation with secondary antibodies is not necessary. The Anti-His-HRP (C-term.) shows even higher sensitivity and specificity for C-terminal His-tagged proteins than the Anti-His-HRP. The latter detects both N- and C-terminal His-tagged proteins.

Antibody specifications
Target:
His epitope (HHHHHH)
Isotype Anti-His antibody: mouse IgG2b
Isotype Anti-His antibody (C-terminal): mouse IgG1
Columns
For µMACS™ Isolation Kit: µ Column, upscale with M Column

For MultiMACS™ Isolation Kits: Multi-8 or Multi-96 Columns
Further information
Protein isolation flyer.pdf
[PDF; 814,4 KB]
 
Figure 1
Purification of a 30 kDa histidine-tagged protein from prokaryotic cell lysate with ProCatch His Resin. Marker (lane 1), lysate from bacterial extract (2), the flow-through (3), and the eluate (4) were separated by SDS-PAGE and Coomassie stained.
Figure 2
Intracellular labeling and FACS analysis of mPD1-His expressing CHO cells. CHO cells were transiently transfected with a mPD1-His encoding vector and analysed by flow cytometry. CHO cells were fixed, permeabilized and labeled with Anti-His-FITC (B) or Mouse IgG1-FITC isotype control (A), respectively.
A
B
Miltenyi Biotec
Toolbox
Customer login
Contact
Newsletter
Help
Share
Print this page
My favorites
Your current favorite list is empty.
Products
μMACS His Isolation Kit
Tag specification
- for 40 isolations
Components
- 2 mL μMACS Anti-His MicroBeads
- 2×50 mL Lysis Buffer
- 50 mL Wash Buffer 1
- 5 mL Wash Buffer 2
- 5 mL Elution Buffer
Download datasheet
130-091-124
Qty.:
 

μMACS Anti-His Starting Kit
- for 40 isolations
Components
- 1 μMACS His Isolation Kit
- 1 μMACS Separation Unit
- 1 MACS MultiStand
- 2×20 μ Columns
130-091-285
Qty.:
 

MultiMACS His Isolation Kit (12×8)
- for 96 isolations
Components
- 3x2 mL µMACS Anti-His MicroBeads
- 1x50 mL Equilibration Buffer
- 1x Multi-8 Column Box (12x8)
Download datasheet
130-094-258
Qty.:
 

MultiMACS His Isolation Kit (4×96)
- for 384 isolations
Components
- 5x4.6 mL µMACS Anti-His MicroBeads
- 1x50 mL Equilibration Buffer
- 1x Multi-96 Column Box (4x96)
Download datasheet
130-094-259
Qty.:
 

Anti-His-FITC
Product Specifications
- 1 mL
- for up to 100 tests
Download datasheet
130-092-675
Qty.:
 

Anti-His-PE
Product Specifications
- 1 mL
- for up to 100 tests
Download datasheet
130-092-691
Qty.:
 

Anti-His-Biotin
Product Specifications
- 1 mL
- for up to 100 tests
Download datasheet
130-092-692
Qty.:
 

Anti-His-HRP
Product Specifications
- 100 µL
Download datasheet
130-092-785
Qty.:
 

Anti-His-HRP (C-term.)
Product Specifications
- 100 µL
Download datasheet
130-092-783
Qty.:
 

ProCatch His Resin
- 10 mL
Download datasheet
130-092-184
Qty.:
 

- 25 mL
Download datasheet
130-092-183
Qty.:
 

- 100 mL
Download datasheet
130-092-182
Qty.:
 

Related products
ProCatch His Resin
Isolation and analysis of c-myc-tagged proteins
Isolation and analysis of GFP-tagged proteins
Isolation and analysis of GST-tagged proteins
Isolation and analysis of HA-tagged proteins
μMACS™ and MultiMACS™ Protein A or Protein G Kits
μMACS™ and MultiMACS™ Streptavidin Kits
MACSelect™ - Transfected Cell Selection Kits
References
1. Dufourcq et al. (2008) American J. Pathology 172(1): 37-49.
Miltenyi Biotec
Miltenyi Biotec
© 1999 - 2010 Miltenyi Biotec ¦ Terms & conditions ¦ Legal notes
Miltenyi Biotec website