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  1. Link

    6x-his-tag-antibody-ab9108.pdf

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Tags & Cell Markers Epitope Tags His Tag
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Anti-6X His tag® antibody (ab9108)

  • Datasheet
  • SDS
Reviews (25)Q&A (4)References (124)

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Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody - ChIP Grade (ab9108)
  • ChIP - Anti-6X His tag® antibody - ChIP Grade (ab9108)
  • Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)
  • Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)

Key features and details

  • Rabbit polyclonal to 6X His tag®
  • Suitable for: WB, ICC/IF, ELISA, ChIP
  • Reacts with: Species independent
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-6X His tag® antibody
    See all 6X His tag® primary antibodies
  • Description

    Rabbit polyclonal to 6X His tag®
  • Host species

    Rabbit
  • Specificity

    ELISA: Antibody specificity was verified by ELISA against peptide conjugated to BSA (HHHHHH-BSA). A 1:25,000 dilution of the antibody gave an OD=1.0 in a 15 minute reaction using HRP-conjugated Goat anti-Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run.
  • Tested applications

    Suitable for: WB, ICC/IF, ELISA, ChIPmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide corresponding to 6X His tag® conjugated to keyhole limpet haemocyanin.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • Purchase matching WB positive control:Recombinant ENO1 protein
  • General notes

    This antibody recognizes both C-terminal and N-terminal tags.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    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: 0.1% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Epitope Tags
    • His Tag

Associated products

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab9108 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
WB (14)
1/1000.
ICC/IF (2)
1/100 - 1/500.
ELISA (2)
1/1000.
ChIP
Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads.
Notes
WB
1/1000.
ICC/IF
1/100 - 1/500.
ELISA
1/1000.
ChIP
Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody - ChIP Grade (ab9108)
    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody - ChIP Grade (ab9108)This image is courtesy of Vladimir Milenkovic

    ab9108 staining CACNB3 with His tag expressed in HEK293 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% goat serum, 0.1% TX100 and PBS) for 16 hours at 4°C. An  Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/750).

    See Abreview

  • ChIP - Anti-6X His tag® antibody - ChIP Grade (ab9108)
    ChIP - Anti-6X His tag® antibody - ChIP Grade (ab9108)
    A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a Myc or His - tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach).
  • Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)
    Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)This image is courtesy of an anonymous Abreview
    All lanes : Anti-6X His tag® antibody (ab9108) at 1/4000 dilution

    Lane 1 : HeLa whole cell lysate - transfected with empty vector
    Lane 2 : HeLa whole cell lysate - transfected with His-tagged gene

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 35 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds

    See Abreview

  • Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)
    Western blot - Anti-6X His tag® antibody - ChIP Grade (ab9108)
    Use of ab9108 in western blotting at 1:1000 against Epitope Tag Positive Control lysate (ab4524).
    Lane 1: 2 µl lysate
    Lane 2: 1 µl lysate
    Lane 3: 0.5 µl lysate
    Lane 4: 0.25 µl lysate Use of ab9108 in western blotting at 1:1000 against E.coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 11 different epitope tag (ab5395). Lane 1: 2 µl lysate Lane 2: 1 µl lysate Lane 3: 0.5 µl lysate Lane 4: 0.25 µl lysate

Protocols

  • ChIP protocols
  • Flow cytometry 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 (124)

Publishing research using ab9108? Please let us know so that we can cite the reference in this datasheet.

ab9108 has been referenced in 124 publications.

  • Liu L  et al. An RFC4/Notch1 signaling feedback loop promotes NSCLC metastasis and stemness. Nat Commun 12:2693 (2021). PubMed: 33976158
  • Hsiue EH  et al. Targeting a neoantigen derived from a common TP53 mutation. Science 371:N/A (2021). PubMed: 33649166
  • Pekas NJ  et al. Design and Development of a Behaviorally Active Recombinant Neurotrophic Factor. Drug Des Devel Ther 14:5393-5403 (2020). PubMed: 33304094
  • Wohlfromm F  et al. Interplay Between Mitophagy and Apoptosis Defines a Cell Fate Upon Co-treatment of Breast Cancer Cells With a Recombinant Fragment of Human ?-Casein and Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand. Front Cell Dev Biol 8:617762 (2020). PubMed: 33537307
  • Yang C  et al. HY5-HDA9 Module Transcriptionally Regulates Plant Autophagy in Response to Light-to-Dark Conversion and Nitrogen Starvation. Mol Plant 13:515-531 (2020). PubMed: 32087368
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 14 Abreviews or Q&A

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
African Green Monkey Recombinant protein (COS 7 cells)
Loading amount
10 µg
Specification
COS 7 cells
Treatment
Homogenised
Gel Running Conditions
Reduced Denaturing (12% tris glycine)
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Mar 27 2013

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Escherichia coli Cell lysate - whole cell (membrane pellet homogenised)
Loading amount
10 µg
Specification
membrane pellet homogenised
Gel Running Conditions
Reduced Denaturing (10-20% Tricine)
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Mar 12 2013

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma cells)
Gel Running Conditions
Reduced Denaturing (4 - 12%)
Loading amount
50000 cells
Treatment
Transfected with His Tagged Protein
Specification
Glioblastoma cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted May 20 2019

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (HeLa)
Specification
HeLa
Treatment
transfected with His-tagged gene
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted Mar 29 2014

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Recombinant protein (HEK Cell supernatant)
Loading amount
40 µg
Specification
HEK Cell supernatant
Gel Running Conditions
Non-reduced Denaturing (15)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Sep 13 2012

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Loading amount
20 µg
Specification
HEK293 cells
Gel Running Conditions
Non-reduced Denaturing (gel 12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Abcam user community

Verified customer

Submitted Nov 11 2011

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (His-tagged purified protein (histone H2B))
Loading amount
0.5 µg
Specification
His-tagged purified protein (histone H2B)
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Read More

Dr. Efrat Shema

Verified customer

Submitted Aug 11 2011

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
20 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

kang sik seo

Verified customer

Submitted Jan 15 2010

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Recombinant protein (TNT system, in vitro translation)
Loading amount
2 µg
Specification
TNT system, in vitro translation
Gel Running Conditions
Non-reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Chien Hsin Lee

Verified customer

Submitted Sep 03 2009

Western blot abreview for Anti-6X His tag® antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells transfected with CACNB3-His)
Loading amount
25 µg
Specification
HEK 293 cells transfected with CACNB3-His
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

HERR DR. Vladimir Milenkovic

Verified customer

Submitted Jun 03 2008

1-10 of 14 Abreviews or Q&A

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