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Western blot - 6X His tag® antibody [4D11] (ab5000)
![Western blot - 6X His tag® antibody [4D11] (ab5000)](/ps/datasheet/images/5/ab5000/6X-His-tag-Primary-antibodies-ab5000-4.jpg)
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Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000)
![Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000)](/ps/datasheet/images/5/ab5000/6X-His-tag-Primary-antibodies-ab5000-5.jpg)
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Immunocytochemistry/ Immunofluorescence - 6X His tag® antibody [4D11] (ab5000)
![Immunocytochemistry/ Immunofluorescence - 6X His tag® antibody [4D11] (ab5000)](/ps/datasheet/images/5/ab5000/6X-His-tag-Primary-antibodies-ab5000-6.jpg)
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Product Name
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6X His tag® antibody [4D11]
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See all 6X His tag antibodies (33)...
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Product type
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Primary antibodies
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Description
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Mouse monoclonal [4D11] to 6X His tag®
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Immunogen
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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'-GCGAGTA AAGCTAAGCT TCCAGCTATT CCACCTGC-3 the amplified DNA sequence was cloned in the vector pDS56/RBSII,6xHis (Hochuli et al. 1988).
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Specificity
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This 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.
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Epitope
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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).
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Tested applications
(see key)
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AP, Dot, ELISA, ICC/IF, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
AP: Use at an assay dependent dilution.
Dot: Use at an assay dependent dilution.
ELISA: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent concentration.
IP: Use at an assay dependent concentration (Abreview).
WB: 1/1000.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Depends upon the localization of the parent protein tagged with hexahistidine.
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Research areas
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Tags & Cell Markers >> Epitope Tags >> His Tag
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Relevance
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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.
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Raised in
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Mouse
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Clonality
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Monoclonal
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Clone number
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4D11
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Myeloma
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NS1
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Isotype
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IgG2a
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Light chain type
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kappa
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Purity
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Ascites
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: Ascites Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Concentration not determined - why is this?
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Storage instructions
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Store 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.
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Notes
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HIS-TAG® is a trademark of EMD Biosciences, Inc.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this 6X His tag® antibody [4D11] is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for 6X His tag® antibody [4D11] images, references, products related to ab5000 and other tools.
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6X His tag® antibody [4D11] images:
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 Western blot - 6X His tag® antibody [4D11] (ab5000)
All lanes : 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
HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
See Abreview
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Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000)
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 image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
See Abreview
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Immunocytochemistry/ Immunofluorescence - 6X His tag® antibody [4D11] (ab5000)
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). This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
See Abreview
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References for 6X His tag® antibody [4D11] (ab5000)
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This product has been used in: (two most recent references)
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. PubMed: 19909498
Mitchell LA et al. Investigation of the role of SRC in capacitation-associated tyrosine phosphorylation of human spermatozoa. Mol Hum Reprod 14:235-43 (2008). PubMed: 18245108
See all 7 publication references for this product.
If you publish research using ab5000 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to 6X His tag®
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6X His tag® antibody [4D11] - more information
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Customer reviews (feedback) regarding 6X His tag® antibody [4D11] |
Customer FAQs regarding 6X His tag® antibody [4D11] |
Protocols for 6X His tag® antibody [4D11] |
Price and availability of products related to 6X His tag® antibody [4D11] |
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6X His tag® antibody [4D11] - related products:
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6X His tag® antibody [4D11] - other tools:
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Contact Abcam with a Technical Enquiry about 6X His tag® antibody [4D11] |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Pricing information loading
![Western blot - 6X His tag® antibody [4D11] (ab5000)](/index.html?datasheet=5000#6X-His-tag-Primary-antibodies-ab5000-4.jpg)
![Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000)](/index.html?datasheet=5000#6X-His-tag-Primary-antibodies-ab5000-5.jpg)
![Immunocytochemistry/ Immunofluorescence - 6X His tag® antibody [4D11] (ab5000)](/index.html?datasheet=5000#6X-His-tag-Primary-antibodies-ab5000-6.jpg)
