His Tag Antibody (3D5) [CoraFluor™ 1]

• Reactivity: ET
• Applications: WB, ELISA
• Clone: 3D5
• Clonality: Monoclonal
• Host: Armenian Hamster
• Conjugate: CoraFluor 1

His Tag Antibody (3D5) [CoraFluor™ 1] Summary

• Description: CoraFluor(TM) 1 is a high performance terbium-based TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) or TRF (Time-Resolved Fluorescence) donor for high throughput assay development. CoraFluor(IM) 1 absorbs UV light at approximately 340 nm, and emits at approximately 490 nm, 545 nm, 585 nm and 620 nm. It is compatible with common acceptor dyes that absorb at the emission wavelengths of CoraFluor(TM) 1. CoraFluor(TM) 1 can be used for the development of robust and scalable TR-FRET binding assays such as target engagement, ternary complex, protein-protein interaction and protein quantification assays.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: This His Tag Antibody (3D5) - Chimeric was developed against a purified his-tagged Fab fragment.
• Specificity: This antibody binds to carboxy-terminal oligo-histidine tags (6x His tags) of fusion proteins. It also has affinity for internal His-tags but no affinity for N-terminal His-tags.
• Isotype: IgG Kappa
• Clonality: Monoclonal
• Host: Armenian Hamster
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Western Blot
• Application Notes: Optimal dilution of this antibody should be experimentally determined.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark. Do not freeze.
• Buffer: PBS
• Preservative: No Preservative
• Purity: Protein A purified

Notes

CoraFluor (TM) is a trademark of Bio-Techne Corp. Sold for research purposes only under agreement from Massachusetts General Hospital. US patent 2022/0025254

Alternate Names for His Tag Antibody (3D5) [CoraFluor™ 1]

• 6 His epitope tag
• 6-His Tag
• 6X His Tag
• 6X His
• 6x-His Tag
• 6X-His
• H
• Hexa His tag
• HHHHHH epitope tag
• HHHHHH tag
• His Tag
• HIS
• His6-Tag
• polyhistidine tag
• polyHistidine
• Poly-histidine

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.