His Tag Antibody [DyLight 405]

Product Discontinued

His Tag Antibody [DyLight 405] Summary

• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to the 6X HIS epitope tag (H-H-H-H-H-H) conjugated to KLH using maleimide.
• Specificity: This affinity purified antibody is directed against the 6X HIS epitope tag and is useful in determining its presence in over expressed proteins in various assays. The antibody recognizes the 6X HIS epitope tag (His-His-His-His-His-His) fused to either the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:340000
  • Fluorophore-linked immunosorbent assay 1:100-1:2000
  • Immunohistochemistry 1:1000-1:5000
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Western Blot 1:2000-1:20000

Reactivity Notes

Non-Species Specific

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Sodium Azide
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute with 0.1 ml deionized water (or equivalent) to a final concentration of 1.0 mg/ml.

Notes

Stabilizer: 10 mg/ml Bovine Serum Albumin (BSA)-Immunoglobulin and Protease free Color: Blue Excitation Wavelength: 400 Emission Wavelength: 420 Conjugation Chemistry: N-hydroxysuccinimide (NHS) ester. Dylight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for His Tag Antibody [DyLight 405]

• 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.