Overview

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • PurityIgG fraction
  • Clonality Monoclonal
  • Clone numberHIS.H8
  • IsotypeIgG2b
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab18184 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
ICC/IF 1/200.
ELISA Use at an assay dependent dilution.
Dot Blot Use at an assay dependent dilution.
WB 1/1000.
IP 1/200.
EMSA Use at an assay dependent dilution. PubMed: 20606007

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 antibody
    • Hexa His tag antibody
    • HHHHHH epitope tag antibody
    • HHHHHH tag antibody
    • His tag antibody
    • Polyhistidine Tag antibody
    see all

Anti-6X His tag® [HIS.H8] antibody images

  • Immunofluorescence (red) with ab18184 on His-tag fusion protein in 293 cells.
    Left hand panel: untransfected control
    Right hand panel: transfected.
    Counterstained with DAPI (blue).

  • All lanes : Anti-6X His tag® [HIS.H8] antibody (ab18184) at 1/1000 dilution

    Lane 1 : 293 cells, untransfected control
    Lane 2 : 293 cells transfected with His-tagged vector
  • Anti-6X His tag® [HIS.H8] antibody (ab18184) at 1/5000 dilution + Conditioned medium from transfected HEK293 cells which produce recombinant secretory protein with a 6XHis-Tag at 5 µg

    Secondary
    An HRP-conjugated Horse anti mouse polyclonal at 1/5000 dilution

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab18184 at a 1/1000 dilution staining 6X His tag in transfected human HEK293 cells by Immunocytochemistry/ Immunofluorescence incubated for 12 hours at 22°C. PFA fixed. Permeabilized using 0.1% Triton X-100 in PBS. Blocked using 3% BSA for 1 hour at 22°C. Secondary used at a 1/1000 dilution polyclonal Goat anti-mouse IgG (H+L) conjugated to Alexa Fluor 488 (green). DAPI staining (blue).

    See Abreview

  • Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 at 1/100 dilution.

    Lane 1 : MW Ladder (from top 120, 85, 50, 34, 25, 19 kDa)
    Lanes 2-5 : Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 from mixture of ~50 ng purified His/FLAG tagged PDCD5 and 15 µL RRL in 300 µL buffer.
    Lanes 6-7 : Purified PDCD5 from E. coli with ~60-70 ng protein.

    The lowest band in each lane is ~ 20 kDa and is PDCD5. The upper bands in lanes 2-5 are the heavy chain (~50 kDa, runs as doublet) and light chain (~25 or 26 kDa).

    Western Blot was performed with a mouse anti-FLAG primary antibody at 1/2000 dilution. An IRDye®800CW-conjugated goat anti-mouse IgG was used as a secondary antibody.

    See Abreview

References for Anti-6X His tag® [HIS.H8] antibody (ab18184)

This product has been referenced in:
  • Booy EP  et al. The RNA helicase RHAU (DHX36) suppresses expression of the transcription factor PITX1. Nucleic Acids Res 42:3346-61 (2014). Read more (PubMed: 24369427) »
  • Andresen CA  et al. Protein interaction screening for the ankyrin repeats and suppressor of cytokine signaling (SOCS) box (ASB) family identify Asb11 as a novel endoplasmic reticulum resident ubiquitin ligase. J Biol Chem 289:2043-54 (2014). WB . Read more (PubMed: 24337577) »

See all 23 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 10 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Recombinant protein (E. coli-purified human VCP-6xHis)
Loading amount 0.01 µg
Specification E. coli-purified human VCP-6xHis
Gel Running Conditions Reduced Denaturing (10% Acrylamide)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 21 2013

The main difference between ab18184 and ab5000 is the immunogen used to produce the antibodies. ab18184 was produced against a 6X His synthetic peptide, while ab5000 was raised against a recombinant protein fused to a 6X His tag. Thus, ab18184 was orig...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - other (PDCD5 Purified from E Coli mixed with RRL)
Total protein in input 100 µg
Specification PDCD5 Purified from E Coli mixed with RRL
Immuno-precipitation step Protein A/G
Username

Dr. Chris Tracy

Verified customer

Submitted Dec 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Recombinant protein (recombinant protein his6-tagged)
Loading amount 0.5 µg
Specification recombinant protein his6-tagged
Gel Running Conditions Reduced Denaturing (Bis-Tris 10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 03 2012

Thank you for contacting us.

I can confirm that all four products, the ab5000, ab15149, ab18184 and ab81663 detect proteins, which contain either an N or C-terminal HHHHHHGS (6x) epitope tag.

Important aspects for his tag recognition ...

Read More

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 30 µg
Specification HEK293
Gel Running Conditions Reduced Denaturing (13)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jul 12 2012

Thank you for contacting Abcam.

The western blot images that are on the webpage for ab18184 were submitted by customers via our Abreviews programand so all of the information we have regarding the protocol they used is available online:
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"