Anti-6X His tag® antibody [4D11] (ab5000)

Overview

• Product nameAnti-6X His tag® antibody [4D11]See all 6X His tag® primary antibodies ...
• Description
  Mouse 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
• Immunogen

  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.

Properties

• FormLiquid
• 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
• MyelomaNS1
• IsotypeIgG2a
• Light chain typekappa
• Research Areas
  • Tags & Cell Markers
  • Epitope Tags
  • His Tag

Applications

Target

• 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.

• Alternative names
  6 His epitope tag antibodyHexa His tag antibodyHHHHHH epitope tag antibody

  HHHHHH tag antibodyHis tag antibody

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Anti-6X His tag® antibody [4D11] images

• 

  Western blot - 6X His tag® antibody [4D11] (ab5000)
  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)
  
  Secondary
  HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution

  This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

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• 

  Immunoprecipitation - 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

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• 

  Western blot - Anti-6X His tag® antibody [4D11] (ab5000)
  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 - 6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
  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).

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