Key features and details
- Mouse monoclonal [4D11] to 6X His tag®
- Suitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cyt
- Reacts with: Species independent
- Isotype: IgG2a
Product nameAnti-6X His tag® antibody [4D11]
See all 6X His tag® primary antibodies
DescriptionMouse monoclonal [4D11] to 6X His tag®
Tested applicationsSuitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cytmore details
Species reactivityReacts with: Species independent
corresponding to 6X His tag®.
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.|
|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
ab5000 has been referenced in 22 publications.
- Lv Z et al. CD19-targeting fusion protein combined with PD1 antibody enhances anti-tumor immunity in mouse models. Oncoimmunology 9:1747688 (2020). PubMed: 32363119
- Chen X et al. A switch in the poly(dC)/RmlB complex regulates bacterial persister formation. Nat Commun 10:27 (2019). PubMed: 30604752
- Herrera T EA et al. GlSir2.1 of Giardia lamblia is a NAD+-dependent cytoplasmic deacetylase. Heliyon 5:e01520 (2019). PubMed: 31025022
- Wang C et al. Lysyl Oxidase-Like Protein 2 Promotes Tumor Lymphangiogenesis and Lymph Node Metastasis in Breast Cancer. Neoplasia 21:413-427 (2019). PubMed: 30925417
- Panagiotakos G et al. Aberrant calcium channel splicing drives defects in cortical differentiation in Timothy syndrome. Elife 8:N/A (2019). PubMed: 31868578