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Anti-6X His tag® antibody [4D11] (ab5000)

Reviews (4)Q&A (6)References (24)
Western blot - Anti-6X His tag® antibody [4D11] (ab5000)
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [4D11] (ab5000)
  • Western blot - Anti-6X His tag® antibody [4D11] (ab5000)
  • Immunoprecipitation - Anti-6X His tag® antibody [4D11] (ab5000)

Key features and details

  • Mouse monoclonal [4D11] to 6X His tag®
  • Suitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cyt
  • Reacts with: Species independent
  • Isotype: IgG2a

Conjugates logo Related conjugates and formulations

HRP

Overview

  • Product name

    Anti-6X His tag® antibody [4D11]
    See all 6X His tag® primary antibodies

  • Description

    Mouse monoclonal [4D11] to 6X His tag®

  • Host species

    Mouse

  • Tested applications

    Suitable for: Dot blot, ELISA, WB, IP, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Species independent

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Epitope

    This antibody was shown to recognize an epitope (HHHHHHGS) encoded by the polylinker of several expression vectors (including pDS:RBSII, pQE-30, and pQE-50).

  • Positive control

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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). Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 6.97% L-Arginine, PBS
  • Concentration information loading...

  • Purity

    Protein G purified

  • Clonality

    Monoclonal

  • Clone number

    4D11

  • Myeloma

    NS1

  • Isotype

    IgG2a

  • Light chain type

    kappa

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab5000 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
AP
Use at an assay dependent concentration.
Dot blot
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
WB (2)
1/1000.
IP (1)
Use at an assay dependent concentration. See Abreview.
ICC/IF (1)
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 
Notes
AP
Use at an assay dependent concentration.
Dot blot
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
WB
1/1000.
IP
Use at an assay dependent concentration. See Abreview.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, 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. 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

  • Western blot - Anti-6X His tag® antibody [4D11] (ab5000)
    Western blot - Anti-6X His tag® antibody [4D11] (ab5000)
    Anti-6X His tag® antibody [4D11] (ab5000) at 1/1000 dilution + 6X His Tagged Protein at 1 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 30 seconds
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [4D11] (ab5000)
    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
    ab5000 staining Human HEK 293 cells transfected with CACNB3-His by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% serum for 20 minutes at 25°C. The primary antibody (1/250 in 1% goat serum, 0.1% TX100, 1X PBS) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat anti-mouse antibody (1/500) was used as the secondary.  DAPI was used for nuclei staining (blue).

    See Abreview

  • Western blot - Anti-6X His tag® antibody [4D11] (ab5000)
    Western blot - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
    All lanes : Anti-6X His tag® antibody [4D11] (ab5000) at 1/2000 dilution

    Lane 1 : Lysates prepared from HEK-293 cells
    Lane 2 : Lysates prepared from HEK-293 cells transfected with Beta3-His (CACNB3)

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

    See Abreview

  • Immunoprecipitation - Anti-6X His tag® antibody [4D11] (ab5000)
    Immunoprecipitation - Anti-6X His tag® antibody [4D11] (ab5000)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

    ab5000 Immunoprecipitate in human HEK293 whole cell lysate. 25µg of cell lysate incubated with primary antibody (undiluted) and matrix (Protein G) for 16 hours at 4°C. For western blotting an undiluted HRP-Goat anti-rabbit IgG polyclonal was used. Beta3-His fusion protein was immunoprecipitated using anti His Ab, and specific beta3a band (56kDa) was detected using anti beta3 Ab

    Lane 1. Lysate of HEK293 cells expressing beta3-His (CACNB3)

    Lane 2. IP with anti His Ab

    Lane 3. Non bound fraction

    See Abreview

References (24)

Publishing research using ab5000? Please let us know so that we can cite the reference in this datasheet.

ab5000 has been referenced in 24 publications.

  • Lv Z  et al. CD19-targeting fusion protein combined with PD1 antibody enhances anti-tumor immunity in mouse models. Oncoimmunology 9:1747688 (2020). PubMed: 32363119
  • Li Y  et al. S100A10 Accelerates Aerobic Glycolysis and Malignant Growth by Activating mTOR-Signaling Pathway in Gastric Cancer. Front Cell Dev Biol 8:559486 (2020). PubMed: 33324631
  • Stephens AD  et al. Isolation and Imaging of His- and RFP-tagged Amyloid-like Proteins from Caenorhabditis elegans by TEM and SIM. Bio Protoc 9:e3408 (2019). PubMed: 33654909
  • Chen X  et al. A switch in the poly(dC)/RmlB complex regulates bacterial persister formation. Nat Commun 10:27 (2019). PubMed: 30604752
  • Herrera T EA  et al. GlSir2.1 of Giardia lamblia is a NAD+-dependent cytoplasmic deacetylase. Heliyon 5:e01520 (2019). PubMed: 31025022
View all Publications for this product

Customer reviews and Q&As

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

1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-6X His tag® antibody [4D11]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells transfected with Beta3-His (CACNB3))
Loading amount
25 µg
Specification
HEK 293 cells transfected with Beta3-His (CACNB3)
Gel Running Conditions
Reduced Denaturing (10% SDS)
Blocking step
Milk as blocking agent for 20 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.

HERR DR. Vladimir Milenkovic

Verified customer

Submitted Nov 20 2009

Western blot abreview for Anti-6X His tag® antibody [4D11]

 Good
Abreviews
Application
Western blot
Sample
Human Recombinant protein (recombinant protein (human))
Loading amount
20 µg
Specification
recombinant protein (human)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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 Nov 27 2006

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