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All lanes: Anti-HIS Tag at 1:2000 dilution + Recombinant protein lysate Lysates/proteins at 20 µg per lane. Secondary: Goat Anti-Mouse IgG, (H+L), Peroxidase conjugated (ASP1613) at 1/8000 dilution. Observed band size: 42 KDa Blocking/Dilution buffer: 5% NFDM/TBST.
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Anti-HIS Tag at 1:8000 dilution + 12-tag lysate Lysates/proteins at 20ng per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 48 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
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Western blot analysis of lysates from 12tag protein , This demonstrates the His tagged antibody detected the His tagged protein (arrow).
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Anti-HIS Tag at 1:2000 dilution + 12 tag recombinant protein lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 45-50 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
- SPECIFICATION
- CITATIONS: 51
- PROTOCOLS
- BACKGROUND
Application 
| WB, E |
|---|---|
| Reactivity | Human |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | Mouse IgG1 |
| Clone/Animal Names | 6AT18 |
| Target/Specificity | Purified recombinant HIS-tagged fusion protein and poly-HIS peptide were used to produced this monoclonal antibody. |
|---|---|
| Dilution | WB~~1:2000 |
| Format | Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein G column, followed by dialysis against PBS. |
| Storage | Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. |
| Precautions | HIS Tag Antibody is for research use only and not for use in diagnostic or therapeutic procedures. |
Provided below are standard protocols that you may find useful for product applications.
Background
Epitope tags consisting of short sequences recognized by well-characterized monoclonal antibodies have been widely used in the study of protein expression in various systems. The 6xHIS tag (HHHHHH), recognized by the monoclonal antibody clone 6AT18 provides an established example of this application. 6xHIS-tagged fusion proteins are easily purified from cell lysates by affinity chromatography using Nickel-Sepharose resin. Abgent's anti-6xHIS monoclonal antibody provides a simple solution to detect the expression of HIS-tagged fusion proteins in cells.
References
Hochuli E, Doebeli H, and Schacher A. New metal chelate absorbent selective for proteins and peptides containing neighboring histidine residues. J. Chromatogr. 1987;411:177-184.
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