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

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Overview

Product name

Anti-6X His tag® antibody [4D11]
See all 6X His tag® products (38) ...

Description

Mouse monoclonal [4D11] to 6X His tag®

Specificity

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.

Tested applications

Dot Blot, ELISA, WB, IP, ICC/IFmore 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).

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).

General notes

HIS-TAG® is a trademark of EMD Biosciences, Inc.

Properties

Form

Liquid

Storage instructions

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.

Storage buffer

Preservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4

Concentration

Concentration information loading...

Purity

IgG fraction

Clonality

Monoclonal

Clone number

4D11

Myeloma

NS1

Isotype

IgG2a

Light chain type

kappa

Research areas

Tags & Cell Markers >> Epitope Tags >> His Tag

  • Western blot - 6X His tag® antibody [4D11] (ab5000)Western blot - 6X His tag® antibody [4D11] (ab5000) image (enlarge)

  • Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000)Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000) image (enlarge)

  • Western blot - Anti-6X His tag® antibody [4D11] (ab5000)Western blot - Anti-6X His tag® antibody [4D11] (ab5000) image (enlarge)

Applications

Show applications key

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.

Target

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
  • HHHHHH tag antibody
  • His tag antibody
see all

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

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

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

See Abreview

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

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

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

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)

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

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

This product has been referenced in:

  • Liu Cet 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) »
  • Mitchell LAet al. Investigation of the role of SRC in capacitation-associated tyrosine phosphorylation of human spermatozoa. Mol Hum Reprod 14:235-43 (2008).Read more (PubMed: 18245108) »

See all 8 publications for this product

Publishing research using ab5000? Please let us know so that we can cite the reference in this datasheet

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"