Overview

  • Product name
    Anti-6X His tag® antibody [HIS.H8]
    See all 6X His tag® primary antibodies
  • Description
    Mouse monoclonal [HIS.H8] to 6X His tag®
  • Host species
    Mouse
  • Specificity
    Recognizes His-tagged recombinant proteins or His-tagged proteins overexpressed in cells.
  • Tested applications
    Suitable for: ICC/IF, ELISA, Dot blot, WB, IP, EMSAmore details
  • Immunogen

    6x His synthetic peptide.

  • Positive control
  • General notes

     

    This product was changed from ascites to tissue culture supernatant on 5th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

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

     

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
    Protein A purified
  • Purification notes
    Purified from TCS by 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

  • 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

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

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

    Performed under reducing conditions.

    Exposure time: 5 minutes

    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 1/5000 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
    All lanes : HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5 minutes


    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

References

This product has been referenced in:
  • Deng L  et al. Double-layered protein nanoparticles induce broad protection against divergent influenza A viruses. Nat Commun 9:359 (2018). Read more (PubMed: 29367723) »
  • Ito N  et al. Ribosome Incorporation into Somatic Cells Promotes Lineage Transdifferentiation towards Multipotency. Sci Rep 8:1634 (2018). Read more (PubMed: 29374279) »
See all 111 Publications for this product

Customer reviews and Q&As

1-10 of 37 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
HEK293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jun 28 2018

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

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

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

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

Abcam user community

Verified customer

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

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

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

Abcam user community

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

Miss. Rachel Loh

Verified customer

Submitted Jan 15 2015

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

Abcam user community

Verified customer

Submitted Aug 04 2014

1-10 of 37 Abreviews or Q&A

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