His Tag Antibody (AD1.1.10) [DyLight 650] Summary
| Immunogen |
This His Tag Antibody (AD1.1.10) - Non-Recombinant Monoclonal was developed against PAX6 transcription factor linked to histidine tag. |
| Isotype |
IgG1 |
| Clonality |
Monoclonal |
| Host |
Mouse |
| Purity |
Protein G purified |
| Innovator's Reward |
Test in a species/application not listed above to receive a full credit towards a future purchase. |
Applications/Dilutions
| Dilutions |
- ELISA
- Flow Cytometry
- Immunohistochemistry
- Immunohistochemistry-Frozen
- Immunoprecipitation
- In vitro assay
- Simple Western
- Western Blot
|
| Application Notes |
Optimal dilution of this antibody should be experimentally determined. |
Reactivity Notes
Packaging, Storage & Formulations
| Storage |
Store at 4C in the dark. |
| Buffer |
50mM Sodium Borate |
| Preservative |
0.05% Sodium Azide |
| Purity |
Protein G purified |
Notes
DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.
Alternate Names for His Tag Antibody (AD1.1.10) [DyLight 650]
Background
A his-tag (also known as histidine tag or polyhistidine tag) is a common epitope tag that typically consists of at least 6 histidine residues fused to either the carboxyl (C-) or amino (N-) terminus of a targeted recombinant protein to facilitate its purification and detection (1). The most common his-tag is the hexahistidine (His6/6-His) tag which has a theoretical molecular weight of 0.8kda (1). The histidine residues readily interact with transition metal ions such as Co2+, Ni2+, Cu2+, and Zn2+, making immobilized metal-affinity chromatography (IMAC) the preferred technique for his-tag purification (1, 2). Metal ions are immobilized and bound to by His-tags in the IMAC column via the histidine imidazole ring. The tagged protein can be eluted off the column by washing with buffers containing a low concentration of imidazole (1, 2). Due to its relatively small size, low immunogenicity, versatility under denaturing conditions, and minimal interference with the structure and function of proteins, the his-tag is one of the most widely used tags for protein purification (1-3).
References
1. Malhotra, A. (2009). Tagging for protein expression. Methods in Enzymology, Guide to Protein Purification, 2nd Edition, 463, 239-258. https://doi.org/10.1016/s0076-6879(09)63016-0
2. Terpe, K. (2003). Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523-533. https://doi.org/10.1007/s00253-002-1158-6
3. Booth, W. T., Schlachter, C. R., Pote, S., Ussin, N., Mank, N. J., Klapper, V., ... Chruszcz, M. (2018). Impact of an N-terminal polyhistidine tag on protein thermal stability. ACS Omega, 3(1), 760-768. https://doi.org/10.1021/acsomega.7b01598
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are
guaranteed for 1 year from date of receipt.
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Product General Protocols
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Video Protocols
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Secondary Antibodies
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Isotype Controls
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Blogs on His Tag
![SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.](https://images.novusbio.com/images/Mouse-IgG1-Isotype-Control-MG1-SDS-Page-NBP1-97005-img0002.jpg "SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.")