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, IHC-P, ICC/IF, ELISA, ChIP, Flow Cytmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Synthetic peptide: HHHHHH conjugated to KLH.

  • Positive control
  • 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
    Preservative: 0.1% Sodium azide
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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 Use at an assay dependent concentration.
WB 1/1000.
IHC-P Use at an assay dependent concentration. PubMed: 21483675
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

  • 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

  • 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).
  • 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

  • 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

References

This product has been referenced in:
  • Diakonov EE  et al. Evidences against vesicle-dependent trafficking and involvement of extracellular proteasomes into cell-to-cell communications. Biochem Biophys Res Commun 508:368-373 (2019). Read more (PubMed: 30503341) »
  • Müskens FM  et al. Design, Synthesis, and Evaluation of a Diazirine Photoaffinity Probe for Ligand-Based Receptor Capture Targeting G Protein-Coupled Receptors. Mol Pharmacol 95:196-209 (2019). Read more (PubMed: 30514721) »
See all 81 Publications for this product

Customer reviews and Q&As

1-10 of 29 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Mar 27 2013

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

Abcam user community

Verified customer

Submitted Mar 12 2013

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

Abcam user community

Verified customer

Submitted Feb 01 2013

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

Abcam user community

Verified customer

Submitted May 20 2019

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

Dayalan Sampath

Verified customer

Submitted Dec 05 2018

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)

Abcam user community

Verified customer

Submitted Oct 21 2016

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

Abcam user community

Verified customer

Submitted Apr 18 2014

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

Abcam user community

Verified customer

Submitted Mar 29 2014

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

Abcam user community

Verified customer

Submitted Sep 13 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

1-10 of 29 Abreviews or Q&A

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