His Tag Antibody (RM146) [DyLight 650] Summary
| Additional Information |
Recombinant Monoclonal Antibody |
| Immunogen |
This His Tag Antibody (RM146) was developed against a mixture of a peptide with 6xHis-Tag at the N-terminus and another peptide with 6xHis-Tag at the C-terminus. |
| Specificity |
This antibody reacts to recombinant proteins containing the 6xHis-Tag or 10xHis-Tag fused to either the amino or carboxy terminus. No cross reactivity with other endogenous protein in mammalian or bacteria cells. |
| Isotype |
IgG |
| Clonality |
Monoclonal |
| Host |
Rabbit |
| Purity |
Protein A purified |
| Innovator's Reward |
Test in a species/application not listed above to receive a full credit towards a future purchase. |
Applications/Dilutions
| Dilutions |
- Flow Cytometry
- Immunocytochemistry/ Immunofluorescence
- Immunohistochemistry
- Immunohistochemistry-Paraffin
- Immunoprecipitation
- Simple Western
- Western Blot
|
| Application Notes |
Optimal dilution of this antibody should be experimentally determined. |
Packaging, Storage & Formulations
| Storage |
Store at 4C in the dark. |
| Buffer |
50mM Sodium Borate |
| Preservative |
0.05% Sodium Azide |
| Purity |
Protein A purified |
Notes
DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.
Alternate Names for His Tag Antibody (RM146) [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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Secondary Antibodies
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Isotype Controls
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Blogs on His Tag
![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
![Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.](https://images.novusbio.com/images/Rabbit--Mouse-IgG-Isotype-Control-Flow-Cytometry-NBP2-24891-img0001.jpg "Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.")