Anti-6X His tag® antibody [4D11] (ab5000)
- 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 applicationsDot Blot, ELISA, WB, IP, ICC/IF more details
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).
- General notesHIS-TAG® is a trademark of EMD Biosciences, Inc.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C (add glycerol to a final volume of 50% for extra stability). Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4
- Concentration information loading...
- PurityIgG fraction
- Clonality Monoclonal
- Clone number4D11
- Light chain typekappa
- Research Areas
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||AP: Use at an assay dependent dilution.|
|Dot Blot||Dot: Use at an assay dependent dilution.|
|ELISA||ELISA: Use at an assay dependent dilution.|
|IP||IP: Use at an assay dependent concentration. See Abreview.|
|ICC/IF||ICC/IF: Use at an assay dependent concentration.|
- 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 antibodyHexa His tag antibodyHHHHHH epitope tag antibody
- HHHHHH tag antibodyHis tag antibody
Anti-6X His tag® antibody [4D11] images
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)
HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution
This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
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
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).
References for Anti-6X His tag® antibody [4D11] (ab5000)
This product has been referenced in:
- Takagi Y et al. Human MTH3 (NUDT18) hydrolyzes the oxidized forms of guanosine and deoxyguanosine diphosphates: A comparison with MTH1 and MTH2. J Biol Chem : (2012). WB . Read more (PubMed: 22556419) »
- Liu C et al. A genome-wide synthetic dosage lethality screen reveals multiple pathways that require the functioning of ubiquitin-binding proteins Rad23 and Dsk2. BMC Biol 7:75 (2009). WB . Read more (PubMed: 19909498) »