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Gene expression profiling products Gene expression profiling products
miRNA expression profiling products miRNA expression profiling products
KIR typing KIR typing
Chimerism analysis Chimerism analysis
Protein isolation and analysis Protein isolation and analysis
  Isolation and analysis of epitope-tagged proteins
  Isolation of interacting molecules with a biotinylated probe
  IP, co-IP and ChIP with Protein A / Protein G MicroBeads
  Native transcription factor isolation
  Large scale recombinant protein isolation
HIV/virus isolation HIV/virus isolation
Organelle research Organelle research
Transfection & transduction Transfection & transduction
MACS® Separators and Columns MACS® Separators and Columns
Genomic services Genomic services
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Isolation and analysis of HA-tagged proteins
Background
Originating from the influenza virus, the hemagglutinin (HA)-epitope (YPYDVPDYA) has become a popular fusion tag for mammalian recombinant protein expression over the last years.

A high-quality monoclonal Anti-HA antibody is offered by Miltenyi Biotec either coupled to µMACS™ MicroBeads for specific isolation of HA-tagged proteins or conjugated to biotin, FITC, PE, or horseradish peroxidase (HRP) for analysis of the recombinant, HA-tagged protein.
Isolation and detection of HA-tagged proteins
Isolation of HA-tagged proteins
The µMACS™ and MultiMACS™ HA Isolation Kits take advantage of the well-established MACS® Technology; thus, allows fast and specific isolation of pure HA-tagged proteins (fig. 1). The advantages of MACS Technology for protein isolation are:

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

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


Detection of HA-tagged proteins
The high-quality monoclonal Anti-HA antibody is available with various conjugates.
The fluorochrome-conjugated (FITC or PE) Anti-HA antibodies enable an immunofluorescence analysis e.g. by fluorescence microscopy (fig. 2), whereas the Anti-HA-HRP antibody was generated for specific and sensitive detection of HA-tagged proteins by Western blot or ELISA analysis. Directly coupled to horseradish peroxidase (HRP), the Anti-HA-HRP antibody simplifies protocols as incubation with secondary antibodies is not necessary.
Varied detection methods are applicable using the biotinylated Anti-HA antibody in combination with appropriate streptavidin conjugates.

Antibody specifications
Target sequence:
YPYDVPDYA
Tag origin: influenca virus hemagglutinin
Isotype: 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 research brochure.pdf
[PDF; 850,1 KB]
 
Figure 1
Sensitive isolation of recombinant fusion protein with μMACS™ Anti-HA MicroBeads. 107 mouse pre-B cells (1881) were transfected with a vector encoding HA-BDCA-2 and proteins isolation was performed using cell populations with either 1% (lane 1, 3) or 10% (2, 4) positively transfected cells. Whole-cell lysates (1, 2) or 20% of the eluate from a protein isolation with Anti-HA MicroBeads (3, 4) were separated by SDS-PAGE, blotted on a membrane and detected by using Anti-HA-HRP.
Figure 2
Immunofluorescence analysis of HA-BDCA-2 expressing RBL-1 cells. RBL-1 (rat basophilic leukemia) cells were transiently transfected with a vector encoding HA-BDCA-2 and analysed by confocal fluorescence microscopy after 24 hours of expression. Cells were first labeled with CD303(BDCA-2)-Biotin, fixed, permeabilized, and then stained with Anti-HA-FITC and Streptavidin-Alexa633. A Fluorescent confocal image of transfected RBL-1 cells. B Overlay of transmission image and fluorescent confocal image of transfected RBL-1 cells.
A
B
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Details
Products
μMACS Anti-HA Starting Kit
- for 40 isolations
Components
- 1 μMACS HA Isolation Kit
- 1 μMACS Separation Unit
- 1 MACS MultiStand
- 2×20 μ Columns
130-091-286
Qty.:
 

μMACS HA Isolation Kit
Tag specification
- for 40 isolations
Components
- 2 mL μMACS Anti-HA MicroBeads
- 2×50 mL Lysis Buffer
- 50 mL Wash Buffer 1
- 5 mL Wash Buffer 2
- 5 mL Elution Buffer
Download datasheet
130-091-122
Qty.:
 

MultiMACS HA Isolation Kit (12×8)
- for 96 isolations
Components
- 3x2 mL µMACS Anti-HA MicroBeads
- 1x50 mL Equilibration Buffer
- 1x Multi-8 Column Box (12x8)
Download datasheet
130-094-255
Qty.:
 

MultiMACS HA Isolation Kit (4×96)
- for 384 isolations
Components
- 5x4.6 mL µMACS Anti-HA MircoBeads
- 1x50 mL Equilibration Buffer
- 1x Multi-96 Column Box (4x96)
Download datasheet
130-094-257
Qty.:
 

Anti-HA-HRP
Specification
- 100 μL
Download datasheet
130-091-972
Qty.:
 

Anti-HA-PE
Specification
- 1 mL
- for up to 100 tests
Download datasheet
130-092-257
Qty.:
 

Anti-HA-FITC
Specification
- 1 mL
- for up to 100 tests
Download datasheet
130-092-256
Qty.:
 

Anti-HA-Biotin
Specification
- 1 mL
- for up to 100 tests
Download datasheet
130-092-258
Qty.:
 

Related products
Isolation and analysis of GFP-tagged proteins
Isolation and analysis of c-myc-tagged proteins
Isolation and analysis of His-tagged proteins
Protein-Protein- / Protein-DNA-Interaction
Isolation and analysis of GST-tagged proteins
μMACS™ and MultiMACS™ Streptavidin Kits
MACSelect™ - Transfected Cell Selection Kits
References
1. Fertey et al. (2010) J. Virol. 84: 9505-9515.
2. Indran et al. (2010) J. Virol. 84: 3162-3177
3. Lattanzi et al. (2010) Hum. Mol. Genet. 19: 2208-2227.
4. Muench et al. (2010) Cancer Res. 70: 6913-6924.
5. Tian et al. (2010) J. Biol. Chem. 285: 23954-23962.
6. Wilton et al. (2010) PNAS 107: 2349-2354.
7. Fujimura et al. (2008) RNA 14: 425-431.
8. Landthaler et al. (2008) RNA 14: 1-17.
9. Pablo-Hernando et al. (2007) Genetics 177: 281-293.
10. Palma et al. (2007) Genes & Dev. 21: 1484-1493.
11. Cole et al. (2006) Nucl. Acids Res. 4: 1281-1292.
12. Gemel et al. (2006) J. Cell. Sci. 119: 2258-2268.
13. Spitzer et al. (2006) Development 133(23): 4679-4689.
14. Weinreich et al. (2006) Blood 108(12): 3713-3721.
15. Richards et al. (2005) Proc. Natl. Acad. Sci. USA 103(5): 1522-1527.
16. Wang et al. (2005) Biochem. Biophys. Res. Com. 333: 1185-1193.
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