Anti-6X His tag® antibody [4D11] (ab5000)
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
Overview
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Product name
Anti-6X His tag® antibody [4D11]
See all 6X His tag® primary antibodies -
Description
Mouse monoclonal [4D11] to 6X His tag® -
Host species
Mouse -
Tested applications
Suitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cytmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
corresponding to 6X His tag®.
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Epitope
This antibody was shown to recognize an epitope (HHHHHHGS) encoded by the polylinker of several expression vectors (including pDS:RBSII, pQE-30, and pQE-50). -
Positive control
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General notes
This antibody clone is manufactured by Abcam.
HIS-TAG® is a trademark of EMD Biosciences, Inc.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 6.97% L-Arginine, PBS -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
4D11 -
Myeloma
NS1 -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
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.
Application | Abreviews | Notes |
---|---|---|
AP | Use at an assay dependent concentration. | |
Dot blot | Use at an assay dependent concentration. | |
ELISA | Use at an assay dependent concentration. | |
WB | 1/1000. | |
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.
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Target
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Relevance
The 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 localization
Depends upon the localization of the parent protein tagged with hexahistidine. -
Alternative names
- 6 His epitope tag antibody
- Hexa His tag antibody
- HHHHHH epitope tag antibody
see all
Images
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Anti-6X His tag® antibody [4D11] (ab5000) at 1/1000 dilution + 6X His Tagged Protein at 1 µg
Secondary
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 -
Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovicab5000 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).
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Western blot - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir MilenkovicAll 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)
Secondary
All lanes : HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution
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Immunoprecipitation - Anti-6X His tag® antibody [4D11] (ab5000)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
Protocols
References (22)
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