Overview

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • Purity
    Ascites
  • Purification notes
    Protein A affinity chromatography
  • Clonality
    Monoclonal
  • Clone number
    HIS.H8
  • Isotype
    IgG2b
  • 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

  • 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
    • Polyhistidine Tag antibody
    see all

Images

  • All lanes : Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/5000 dilution

    Lane 1 : ARPE-19 whole cell lysate - non-transfected
    Lane 2 : ARPE-19 whole cell lysate - transfected with rRab27a-His

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Vladimir Milenkovic

    Blocked with 5% milk for 30 minutes at 25°C.

    Incubated with the primary antibody for 16 hours at 4°C in 1X PBS/Tween, 5% milk.

    See Abreview

  • LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue).

  • 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

  • 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

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

    Lane 1 : 6x His-tagged protein expressed in BL21 (pET15b) at 1 µg
    Lane 2 : 6x His-tagged protein expressed in BL21 (pET15b) at 0.1 µg

    Secondary
    IRDye® 800CW goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution

    Performed under reducing conditions.

    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Rachel Loh

    See Abreview

References

This product has been referenced in:
  • Dickens D  et al. Modulation of LAT1 (SLC7A5) transporter activity and stability by membrane cholesterol. Sci Rep 7:43580 (2017). WB . Read more (PubMed: 28272458) »
  • Wang BD  et al. Alternative splicing promotes tumour aggressiveness and drug resistance in African American prostate cancer. Nat Commun 8:15921 (2017). IP ; Mouse . Read more (PubMed: 28665395) »

See all 68 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (normal fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
100 µg
Treatment
MG132, and UVA
Specification
normal fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 1°C
Username

Abcam user community

Verified customer

Submitted Dec 11 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
10 µg
Immuno-precipitation step
Protein A/G
Specification
293T
Username

Abcam user community

Verified customer

Submitted Oct 16 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela cells)
Permeabilization
Yes - 0.1 % Triton X100
Specification
Hela cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 13 2017

Application
Western blot
Sample
Human Recombinant protein (Recombinant protein)
Gel Running Conditions
Non-reduced Denaturing (12)
Loading amount
0.1 µg
Specification
Recombinant protein
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

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Submitted Oct 13 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Embryonic fibrobalst cells)
Total protein in input
1e+006 cells
Treatment
Influenza virus infection
Immuno-precipitation step
Other - Protein G - Dynabeads
Specification
Embryonic fibrobalst cells
Username

Abcam user community

Verified customer

Submitted Mar 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spinal Cord (Dorsal Horn))
Antigen retrieval step
None
Permeabilization
Yes - 1% Triton, 0.25% Tween in 1XPBS
Specification
Spinal Cord (Dorsal Horn)
Blocking step
Mouse-on-Mouse Block (Vector Labs) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 11 2016

Application
Western blot
Sample
Human Purified protein (S2 cell)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
0.001 µg
Specification
S2 cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted May 27 2015

Application
Western blot
Loading amount
1 µg
Gel Running Conditions
Reduced Denaturing (12% hand cast gel)
Sample
Escherichia coli Recombinant protein (6x His-tagged protein expressed in BL21 (pET15b))
Specification
6x His-tagged protein expressed in BL21 (pET15b)
Blocking step
Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Miss. Rachel Loh

Verified customer

Submitted Jan 15 2015

Application
IHC - Wholemount
Sample
Xenopus tropicalis Embryo (whole embryo)
Specification
whole embryo
Username

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Verified customer

Submitted Aug 04 2014

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

1-10 of 36 Abreviews or Q&A

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