His Tag Antibody (RMH01) - Chimeric

Western Blot: His Tag Antibody (RMH01) [NBP2-61478] - Western blot of 293T cells transfected (+) or untransfected (-) with a DNA construct encoding His-Tag Histone H3 (G34W) protein, using NBP2-61478 at 0.2 ug/mL, followed by a HRP conjugated anti-Human IgG secondary antibody.

Sandwich ELISA: His Tag Antibody (RMH01) - Chimeric - Azide and BSA Free [NBP2-61478] - Sandwich ELISA performed on different amounts of whole lysate of 293T cells transfected with a DNA construct encoding His Tag Histone H3 (K36M) protein, using Clone RMH01 as the capture antibody. Another Histone H3 [monomethyl Lys 36] rabbit monoclonal antibody was used as the detection antibody, followed by an alkaline phosphatase conjugated Rabbit IgG as the Secondary antibody.

ELISA: His Tag Antibody (RMH01) - Chimeric - Azide and BSA Free [NBP2-61478] - A titer ELISA performed on purified His Tag-PCSK9 recombinant protein. A serial dilution of Clone RMH01 was used as the primary antibody. An alkaline phosphatase conjugated anti-Human IgG as the secondary antibody.

• Reactivity: ET
• Applications: WB, Simple Western, ELISA, ELISA
• Clone: RMH01
• Clonality: Monoclonal
• Host: Human
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

His Tag Antibody (RMH01) - Chimeric Summary

• Description: Novus Biologicals Human His Tag Antibody (RMH01) - Chimeric (NBP2-61478) is a recombinant monoclonal antibody validated for use in WB, ELISA and Simple Western. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: This His Tag Antibody (RMH01) 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. This antibody is a Rabbit-Human Chimeric antibody, made by fusion of the antigen binding region (variable domains of the heavy and light chains, VH and VL) of NBP2-61482, with the constant domain of Human IgG1. The antibody is recognized only by an anti-Human IgG secondary antibody.
• Source: HEK293
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Human
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Sandwich ELISA Capture 0.1 - 2 ug/ml
  • Sandwich ELISA Detection 0.01 - 0.5 ug/ml
  • Simple Western
  • Western Blot 0.1 ug/ml - 1 ug/ml
• Application Notes: See Simple Western Antibody Database for Simple Western validation: Tested in Human purified protein, separated by Size

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: 50% Glycerol/PBS, 1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for His Tag Antibody (RMH01) - Chimeric

• 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.

Review for His Tag Antibody (NBP2-61478) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer Simple Western Human 12/04/2018

Summary

• Application: Simple Western
• Sample Tested: Purified protein
• Species: Human
• Lot: R-08-01719