Plasmid vectors for the expression of coding regions of eukaryotic genes in bacterial, insect and mammalian hosts are in common usage; such expression vectors are frequently used to encode hybrid fusion proteins consisting of a eukaryotic target protein and a specialized region designed to aid in the purification and visualization of the target protein. A system that has proven to be very successful relies on the insertion of a six histidine (His6) sequence in the N-terminus of the encoded protein, allowing for efficient coupling to Ni -chelating resins and purification by single step affinity chromatography. This polyhistidine sequence can then be removed by specific cleavage at sites recognized by enzymes such as thrombin or enterokinase, permitting the separation of the target protein from the polyhistidine tag. Visualization of such fusion proteins can be achieved by utilizing antibodies generated against specific peptide sequences downstream from the multiple cloning site.

1. O-GlcNAcylation of SIRT1 enhances its deacetylase activity and promotes cytoprotection under stress.Nat Commun. 2017 Nov 14;8(1):1491

2. Oligomerization-primed coiled-coil domain interaction with Ubc13 confers processivity to TRAF6 ubiquitin ligase activity. Nat Commun. 2017 Oct 9;8(1):814

3. Phosphorylation of SPT5 by CDKD;2 Is Required for VIP5 Recruitment and Normal Flowering in Arabidopsis thaliana. Plant Cell. 2017 Feb;29(2):277-291.

4. Ubiquitylation of MFHAS1 by the ubiquitin ligase praja2 promotes M1 macrophage polarization by activating JNK and p38 pathways. Cell Death Dis. 2017 May 4;8(5):e2763.

5. Red blood cells release microparticles containing human argonaute 2 and miRNAs to target genes of Plasmodium falciparum. Emerg Microbes Infect. 2017 Aug 23;6(8):e75

6. Transmembrane domain is crucial to the subcellular localization and function of Myc target 1. J Cell Mol Med. 2016 Mar;20(3):471-81.

7. Crystal structure of Gib2, a signal-transducing protein scaffold associated with ribosomes in Cryptococcus neoformans. Sci Rep. 2015 Mar 3;5:8688.

8. C-terminal domain of leucyl-tRNA synthetase from pathogenic Candida albicans recognizes both tRNASer and tRNALeu. J Biol Chem. 2016 Feb 12;291(7):3613-25

9. The EZH1–SUZ12 complex positively regulates the transcription of NF-κB target genes through interaction with UXT. J Cell Sci 2016 129: 2343-2353

10. MoDnm1 Dynamin Mediating Peroxisomal and Mitochondrial Fission in Complex with MoFis1 and MoMdv1 Is Important for Development of Functional Appressorium in Magnaporthe oryzae. PLoS Pathog. 2016 Aug 24;12(8):e1005823.

 

This Abmart monoclonal antibody is produced by immunizing mice with a 6×His synthetic peptide (KLH-coupled).

His-Tag (2A8) Mouse mAb detects over-expressed or recombinant proteins containing the 6× His epitope tag.

Store at -20°C. Stable for one year from the date of shipment.

All
 

Mouse IgG1
 

Use an anti-Mouse secondary antibody to detect the 2A8 antibody.

Western blotting            1:5000
Immunofluorescence   1:2000
Immunoprecipitation     1:100
ELISA                           1:2000