Product nameAnti-6X His tag® antibody [4D11]
See all 6X His tag® primary antibodies
DescriptionMouse monoclonal [4D11] to 6X His tag®
SpecificityThis antibody recognises all proteins which contain either an N or C-terminal HHHHHHGS epitope tag. It has been shown by a reviewer to recognise HHHHHH alone.
Tested applicationsSuitable for: Dot blot, ELISA, WB, IP, ICC/IF, Other, Flow Cytmore details
Species reactivityReacts with: Species independent
Recombinant malaria protein TRAP fused at the amino-terminal end with six His tag. The coding sequence of TRAP 1.0 was amplified from P. falciparum genomic DNA (ITO4 isolate) in standard Polymerase Chain Reaction PCR (Saiki et al. 1988) using the primer combination trap 1 5'-AGTTGGATCCAGAGATGTGCAAAACA ATATAGTGG-3' and trap 2 5'-GCGAGTAAAGCTAAGCTTCCAGCTATTCCACCTGC-3', the amplified DNA sequence was cloned in the vector pDS56/RBSII,6xHis (Hochuli et al. 1988).
EpitopeThis antibody was shown to recognize an epitope (HHHHHHGS) encoded by the polylinker of several expression vectors (including pDS:RBSII, pQE-30, and pQE-50).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 6.97% L-Arginine, PBS
Concentration information loading...
Light chain typekappa
Our Abpromise guarantee covers the use of ab5000 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|AP||Use at an assay dependent concentration.|
|Dot blot||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration. See Abreview.|
|ICC/IF||Use at an assay dependent concentration.|
|Other||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
RelevanceThe H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
Cellular localizationDepends upon the localization of the parent protein tagged with hexahistidine.
- 6 His epitope tag antibody
- Hexa His tag antibody
- HHHHHH epitope tag antibody
Anti-6X His tag® antibody [4D11] (ab5000) at 1/1000 dilution + 6X His Tagged Protein at 1 µg
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 30 seconds
ab5000 staining Human HEK 293 cells transfected with CACNB3-His by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% serum for 20 minutes at 25°C. The primary antibody (1/250 in 1% goat serum, 0.1% TX100, 1X PBS) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat anti-mouse antibody (1/500) was used as the secondary. DAPI was used for nuclei staining (blue).
All lanes : Anti-6X His tag® antibody [4D11] (ab5000) at 1/2000 dilution
Lane 1 : Lysates prepared from HEK-293 cells
Lane 2 : Lysates prepared from HEK-293 cells transfected with Beta3-His (CACNB3)
All lanes : HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution
ab5000 Immunoprecipitate in human HEK293 whole cell lysate. 25µg of cell lysate incubated with primary antibody (undiluted) and matrix (Protein G) for 16 hours at 4°C. For western blotting an undiluted HRP-Goat anti-rabbit IgG polyclonal was used. Beta3-His fusion protein was immunoprecipitated using anti His Ab, and specific beta3a band (56kDa) was detected using anti beta3 Ab
Lane 1. Lysate of HEK293 cells expressing beta3-His (CACNB3)
Lane 2. IP with anti His Ab
Lane 3. Non bound fraction
This product has been referenced in:
- Zhang J et al. DAO1 catalyzes temporal and tissue-specific oxidative inactivation of auxin in Arabidopsis thaliana. Proc Natl Acad Sci U S A 113:11010-5 (2016). WB . Read more (PubMed: 27651492) »
- Merriman C et al. Lipid-tuned Zinc Transport Activity of Human ZnT8 Protein Correlates with Risk for Type-2 Diabetes. J Biol Chem 291:26950-26957 (2016). Read more (PubMed: 27875315) »