His Tag Antibody (AD1.1.10) - Azide and BSA Free 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 |
- CyTOF-ready
- ELISA 1:100 - 1:500
- Flow Cytometry 1:10 - 1:50
- Immunohistochemistry 1:100 - 1:300
- Immunohistochemistry-Frozen 1:100 - 1:300
- Immunoprecipitation 1:10 - 1:500
- In vitro assay
- Simple Western 1:250
- Western Blot 1:1000
|
Application Notes |
This antibody is routinely tested in WB using Histidine-tagged recombinant proteins. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. |
Packaging, Storage & Formulations
Storage |
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
Buffer |
PBS |
Preservative |
No Preservative |
Concentration |
1 mg/ml |
Purity |
Protein G purified |
Alternate Names for His Tag Antibody (AD1.1.10) - Azide and BSA Free
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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