6xHistidine Epitope Tag

Principal name

6xHistidine Epitope Tag antibody

Selected pictures

6xHistidine Epitope Tag antibody - AM00195PU-N
AM00195PU-N - Western blot analysis of His in 293 cells transfectedwith His-tagged vector (Lane 2) and Control vector (Lane 1).
6xHistidine Epitope Tag antibody - AM00195PU-N
AM00195PU-N - Immunofluorescence of His-tagged fusion protein expressed in 293 cells. Left Lane: Untransfected control; Right Lane: Transfected cells.

Available reactivities

All Species, Hu (Human)

Available hosts

Mouse, Rabbit, Goat, Chicken

Available applications

Immunocytochemistry/Immunofluorescence (ICC/IF), Immunoprecipitation (IP), Western blot / Immunoblot (WB), Enzyme Immunoassay (E), Paraffin Sections (P), Frozen Sections (C), Flow Cytometry (F), ELISA (capture) (E(capture)), ELISA (detection) (E(detection))

Background of 6xHistidine Epitope Tag antibody

In the last couple of years many peptide sequences/epitopes for the purification of recombinant proteins have been established. These so-called “tags” can be used e.g. to determine the cellular localization or to quantify proteins. The polyhistidine “tag” (His-tag) is the most used affinity epitope for the purification of recombinant proteins [1]. Proteins with a polyhistidine tag (e.g. 6xHis or 8xHis) can be purified in one step using a metal-chelate column (e.g. Ni2+, Zn2+, Cu2+ or Co2+) and imidazole as eluent. This method now is a very attractive system for the purification of larger amounts proteins for structural and functional studies. So far His-tagged proteins were successfully purified from different expression systems like E. coli, yeast, insect cells and plant cells [1,2]. An important requirement beside the efficient and robust purification method is the availability of a fast detection system for checking the purification steps of these His-tagged proteins if no specific antibody is available.

General readings

1. Conrath, K. et al. (2001) Camel single binding-domain antibodies as modular binding units in bispecific and bivalent antibody constructs. J. Biol.Chem. 276 (10): 7346-7350
2. Suen, J.L. et al. (2001) Characterization of self-T-cell response and antigenic determinant of U1A protein with bone marrow-derived dendritic cells in NZB x NZW F1 mice. Immunol. 103: 301-309.
3. Hoffmann, S.C. et al. (2007) Identification of CLEC12B, an inhibitory receptor on myeloid cells. J. Biol. Chem. 282: 22370-22375.
4. Zheng, J. et al. (2007) Serum from mice immunized in the context of Treg inhibition identifies DEK as a neuroblastoma tumor antigen. BMC Immunol. 8: 4.
5. Bahi, A. & Dreyer, J.L. (2008) Overexpression of plasminogen activators in the nucleus accumbens enhances cocaine-, amphetamine- and morphine-induced reward and behavioral sensitization. Genes Brain Behav. 7: 244-256.
6. Wrighton, K.H. et al. (2009) Transforming Growth Factor beta can stimulate Smad1 phosphorylation independently of Bone Morphogenic Protein receptors. J. Biol. Chem. 284: 9755-9763.
7. Diefenbacher, M. et al., The Dsl1 Tethering Complex Actively Participates in Soluble NSF (N-Ethylmaleimide-sensitive Factor) Attachment Protein Receptor (SNARE) Complex Assembly at the Endoplasmic Reticulum in Saccharomyces cerevisiae. J Biol Chem. 286: 25027-25038 (2011).
8. Alvarez, M.M. et al., Specific recognition of influenza A/H1N1/2009 antibodies in human serum: a simple virus-free ELISA method. PLoS One. 5: e10176 (2010).
9. Bahi, A. et al., The role of tissue-type plasminogen activator system in amphetamine-induced conditional place preference extinction and reinstatement. Neuropsychopharmacology 33: 2726-2735 (2008).
10. Gunnarsen, K.S. et al., Periplasmic expression of soluble single chain T cell receptors is rescued by the chaperone FkpA.  BMC Biotechnol. 10: 8 (2010).
11. Hwang, H.Y. et al., Highly specific inhibition of C1q globular-head binding to human IgG: a novel approach to control and regulate the classical complement pathway using an engineered single chain antibody variable fragment. Mol Immunol. 45: 2570-2580 (2008).
12. De Vooght, L. et al., Expression and extracellular release of a functional anti-trypanosome Nanobody® in Sodalis glossinidius, a bacterial symbiont of the tsetse fly. Microb Cell Fact. 11: 23 (2012).

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Antibodies

Catalog No. Host Clone/Iso. Pres.   React. Applications  
R1181

6xHistidine Epitope Tag (HHHHHH) antibody

Figure 1. Anti-6X His epitope tag polyclonal antibody detects His-tagged recombinant proteins by western blot. The blot was blocked with 3% BSA in TBST for 45 min at RT. Antibody was incubated with blot at a 1/1,000 dilution in TBST with 3% BSA for 1 hour at RT. Detection occurred using HRP Goat-anti-Rabbit IgG  diluted 1/80,000 in blocking buffer for 30 min at RT. Lane 1 was loaded with 12-Epitope Tag Protein Marker Lysate which has the His epitope tag incorporated through a C-terminal linkage (~42 kDa). Lane 2 was loaded with His-SUMO-GFP recombinant protein which has the His epitope tag incorporated through an N-terminal linkage (~40 kDa). A 4-20% gradient gel was used to resolve the protein by SDS-PAGE. Proteins were transferred to nitrocellulose using standard methods. Molecular weights were estimated by comparison to standards (lane M). Rabbit Aff - Purified C, E, WB
0.1 mg / €240.00
    Acris Antibodies GmbH
 
18812

6xHistidine Epitope Tag (HHHHHH) antibody

6xHistidine Epitope Tag Goat Aff - Purified C, E, ICC/IF, WB
1 mg / €920.00
    QED Bioscience Inc.
 
18812-01

6xHistidine Epitope Tag (HHHHHH) antibody

6xHistidine Epitope Tag Goat Aff - Purified C, E, ICC/IF, WB
0.1 mg / €290.00
    QED Bioscience Inc.
 
18814

6xHistidine Epitope Tag (HHHHHH) antibody

6xHistidine Epitope Tag Rabbit Aff - Purified C, E, ICC/IF, WB
1 mg / €920.00
    QED Bioscience Inc.
 
18814-01

6xHistidine Epitope Tag (HHHHHH) antibody

6xHistidine Epitope Tag Rabbit Aff - Purified C, E, ICC/IF, WB
0.1 mg / €290.00
    QED Bioscience Inc.
 

5 Item(s)

per page