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  1. Link

    products/primary-antibodies/6x-his-tag-antibody-hish8-ab18184.pdf

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Tags & Cell Markers Epitope Tags His Tag
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Anti-6X His tag® antibody [HIS.H8] (ab18184)

  • Datasheet
  • SDS
Reviews (32)Q&A (10)References (250)

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Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)
  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)
  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)

Key features and details

  • Mouse monoclonal [HIS.H8] to 6X His tag®
  • Suitable for: ICC/IF, ELISA, Dot blot, WB, IP
  • Reacts with: Species independent
  • Isotype: IgG2b

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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.

    ab18184 reacts to recombinant proteins containing the 6X His tag® or 10X His tag® fused to either the amino or carboxy terminus.

  • Tested applications

    Suitable for: ICC/IF, ELISA, Dot blot, WB, IPmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide corresponding to 6X His tag®.

  • Positive control

    • Purchase matching WB positive control:Recombinant ENO1 protein
  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • 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

    • Tags & Cell Markers
    • Epitope Tags
    • His Tag
    • Signal Transduction
    • Antibodies
    • 6x his tag®

Associated products

  • Alternative Versions

    • Biotin Anti-6X His tag® antibody [HIS.H8] (ab173828)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)

Applications

The Abpromise guarantee

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 (3)
1/200.
ELISA (1)
Use at an assay dependent concentration.
Dot blot
Use at an assay dependent concentration.
WB (21)
1/1000.
IP (5)
1/200.
Notes
ICC/IF
1/200.
ELISA
Use at an assay dependent concentration.
Dot blot
Use at an assay dependent concentration.
WB
1/1000.
IP
1/200.

Target

  • Relevance

    The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. His-tags can be fused to the amino- or carboxy- termini of 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 - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Mr Chris Tracy

    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 6-9 : 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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Vladimir Milenkovic
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)

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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Rachel Loh
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an anonymous 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

Protocols

  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (250)

Publishing research using ab18184? Please let us know so that we can cite the reference in this datasheet.

ab18184 has been referenced in 250 publications.

  • Smith EM  et al. The mTOR regulated RNA-binding protein LARP1 requires PABPC1 for guided mRNA interaction. Nucleic Acids Res 49:458-478 (2021). PubMed: 33332560
  • Lim Y  et al. Phosphorylation by Aurora B kinase regulates caspase-2 activity and function. Cell Death Differ 28:349-366 (2021). PubMed: 32811973
  • Wang S  et al. AXL is a candidate receptor for SARS-CoV-2 that promotes infection of pulmonary and bronchial epithelial cells. Cell Res 31:126-140 (2021). PubMed: 33420426
  • Kwak MS  et al. Reactive oxygen species induce Cys106-mediated anti-parallel HMGB1 dimerization that protects against DNA damage. Redox Biol 40:101858 (2021). PubMed: 33461096
  • Calero-Cuenca FJ  et al. Ctdnep1 and Eps8L2 regulate dorsal actin cables for nuclear positioning during cell migration. Curr Biol 31:1521-1530.e8 (2021). PubMed: 33567288
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 42 Abreviews or Q&A

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Recombinant protein (Overexpressed his-tagged protein in a eColi cell c)
Gel Running Conditions
Reduced Denaturing
Loading amount
1 µg
Specification
Overexpressed his-tagged protein in a eColi cell c
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 35°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 13 2020

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (purified protein)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
5 µg
Treatment
fulllength, deletion mutant expression
Specification
purified protein
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 16 2020

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Fish Recombinant protein (sf9)
Gel Running Conditions
Reduced Non-Denaturing (Native) (10% SDS)
Loading amount
2e+007 cells
Specification
sf9
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 02 2020

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Escherichia coli Cell lysate - whole cell (Escherichia coli)
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
30 µg
Specification
Escherichia coli
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Lawrence Welch

Verified customer

Submitted Jun 15 2019

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (human, purified protein)
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
0.25 µg
Specification
human, purified protein
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 20 2019

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 28 2018

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 11 2017

Immunoprecipitation abreview for Anti-6X His tag® antibody [HIS.H8]

Good
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
10 µg
Immuno-precipitation step
Protein A/G
Specification
293T
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 16 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-6X His tag® antibody [HIS.H8]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 13 2017

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 13 2017

1-10 of 42 Abreviews or Q&A

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