His Tag Antibody (3D5) - Chimeric - Azide and BSA Free

Simple Western: His Tag Antibody (3D5) - Chimeric [NBP2-52637] - His Tag Antibody (3D5) [NBP2-52637] - Human purified protein. Simple Western image submitted by a verified customer review.

Western Blot: His Tag Antibody (3D5) - Chimeric - Azide and BSA Free [NBP2-52637] - His tag expressing plasmid transfected (A) and non-transfected (B) HEK293 cells lysate (3g protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with NBP2-52637 at 0.01ug/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

Immunocytochemistry /Immunofluorescence: His Tag Antibody (3D5) - Chimeric - Azide and BSA Free [NBP2-52637] - Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells transfected with His tag expressing plasmid (A) and non-transfected HEK293 cells (B), permeabilized with 0.15% Triton stained with NBP2-52637 (1:200 dilution) for 1h followed by Alexa Fluor 488 secondary antibody (1:1000 dilution), showing cytoplasmic staining. The nuclear stain is DAPI (blue).

• Reactivity: ET
• Applications: WB, Simple Western, ELISA
• Clone: 3D5
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

His Tag Antibody (3D5) - Chimeric - Azide and BSA Free Summary

• Description: Novus Biologicals Rabbit His Tag Antibody (3D5) - Chimeric - Azide and BSA Free (NBP2-52637) 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 (3D5) - Chimeric was developed against 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: Rabbit
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100 - 1:2000
  • Simple Western
  • Western Blot 1:100 - 1:2000
• Application Notes: This is a chimeric rabbit antibody, based on the original mouse IgG2b format, created for improved compatibility with existing reagents, assays and techniques.
  See Simple Western Antibody Database for Simple Western validation: Tested in Human purified protein, separated by Size

Packaging, Storage & Formulations

• Storage: Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.
• Buffer: PBS
• Preservative: 0.02% Proclin 300
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for His Tag Antibody (3D5) - Chimeric - Azide and BSA Free

• 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 (NBP2-52637) (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: T1538A11