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    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 (108)

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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: IP, WB, ICC/IF, ELISA, ChIP, Flow Cyt
  • Reacts with: Species independent
  • Isotype: IgG

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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: IP, WB, ICC/IF, ELISA, ChIP, Flow Cytmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide: HHHHHH conjugated to KLH.

  • Positive control

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

    This antibody recognizes both C-terminal and N-terminal tags. HIS-TAG® is a trademark of EMD Biosciences, Inc.

    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.

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
IP (5)
Use at an assay dependent concentration.
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.
Flow Cyt (1)
Use at an assay dependent concentration.

(see Abreview published on 22nd January, 2007).

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
IP
Use at an assay dependent concentration.
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.
Flow Cyt
Use at an assay dependent concentration.

(see Abreview published on 22nd January, 2007).

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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

  • 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

    • Datasheet
    • SDS
  • References (108)

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

    ab9108 has been referenced in 108 publications.

    • Schommers P  et al. Restriction of HIV-1 Escape by a Highly Broad and Potent Neutralizing Antibody. Cell 180:471-489.e22 (2020). PubMed: 32004464
    • Jarsch IK  et al. A direct role for SNX9 in the biogenesis of filopodia. J Cell Biol 219:N/A (2020). PubMed: 32328641
    • Liu J  et al. Long noncoding RNA AGPG regulates PFKFB3-mediated tumor glycolytic reprogramming. Nat Commun 11:1507 (2020). PubMed: 32198345
    • Tufegdžic Vidakovic A  et al. Regulation of the RNAPII Pool Is Integral to the DNA Damage Response. Cell 180:1245-1261.e21 (2020). PubMed: 32142654
    • Patra D  et al. Site-1 protease ablation in the osterix-lineage in mice results in bone marrow neutrophilia and hematopoietic stem cell alterations. Biol Open 9:N/A (2020). PubMed: 32576566
    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 29 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

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - other (HEK Cell memrane lysate incubated with supernatant)
    Total protein in input
    100000 cells
    Specification
    HEK Cell memrane lysate incubated with supernatant
    Immuno-precipitation step
    Protein G
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 01 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

    Immunohistochemistry (Frozen sections) abreview for Anti-6X His tag® antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain, His-tagged EPO)
    Permeabilization
    Yes - Tween-20
    Specification
    Brain, His-tagged EPO
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C
    Fixative
    Acetone
    Read More

    Dayalan Sampath

    Verified customer

    Submitted Dec 05 2018

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Cell (NIH3T3)
    Specification
    NIH3T3
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Type
    Sandwich (Capture)
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 21 2016

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein G
    Sample
    Human Cell lysate - whole cell (HeLa)
    Specification
    HeLa
    Treatment
    100ug/ml FAC or 100uM DFO for 8hrs
    Total protein in input
    5 µg
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 18 2014

    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

    Question

    I read in the description of one of your anti-6xHIS tag antibodies
    (ab9108) that it was generated using an 6xHistagged version of bovine
    seruml albumine (HHHHHH-BSA). I quote: "ELISA: Antibody specificity was
    verified by ELISA against peptide conjugated to BSA (HHHHHH-BSA)".
    My question to you is: do you sell this his-tagged BSA as well?

    Read More

    Abcam community

    Verified customer

    Asked on May 04 2012

    Answer

    Unfortunately we do not have a His tagged BSA in our catalog and can not offer HIS-tagged BSA for sale as it's just a component within our QC procedures.

    Read More

    Abcam Scientific Support

    Answered on May 04 2012

    1-10 of 29 Abreviews or Q&A

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