Alkaline Phosphatase Anti-6X His tag® antibody [HIS-1] (ab49746)

Reviews (1)Q&A (1)References (3)

Key features and details

  • Alkaline Phosphatase Mouse monoclonal [HIS-1] to 6X His tag®
  • Suitable for: WB, Dot blot, ELISA
  • Reacts with: Species independent
  • Conjugation: Alkaline Phosphatase
  • Isotype: IgG2a

Overview

  • Product name

    Alkaline Phosphatase Anti-6X His tag® antibody [HIS-1]
    See all 6X His tag® primary antibodies

  • Description

    Alkaline Phosphatase Mouse monoclonal [HIS-1] to 6X His tag®

  • Host species

    Mouse

  • Conjugation

    Alkaline Phosphatase

  • Tested applications

    Suitable for: WB, Dot blot, ELISAmore details

  • Species reactivity

    Reacts with: Species independent

  • Immunogen

    Recombinant poly(Histidine)-tagged fusion protein.

  • Positive control

    • Bacteria lysates expressing a recombinant polyHistidine tagged fusion protein and BCIP/NBT substrate.

  • General notes

    HIS-TAG® is a trademark of EMD Biosciences, Inc.

    Monoclonal antibodies reacting specifically with poly-histidine may be useful in various immunotechniques, to identify the expression of a polyhistidine fusion protein in bacteria, bacterial lysates or cells and tissues transfected with a polyHistidinetagged fusion protein expressing vectors.

    The antibody is prepared by conjugation of calf intestinal alkaline phosphatase to mouse monoclonal anti-polyHistidine purified from ascites fluid of the HIS-1 hybridoma

    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). Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 8.00
    Preservative: 0.097% Sodium azide
    Constituents: 0.06% Tris buffered saline, 0.0095% Magnesium chloride, 50% Glycerol, 1% BSA
  • Concentration information loading...

  • Primary antibody notes

    Monoclonal antibodies reacting specifically with poly-histidine may be useful in various immunotechniques, to identify the expression of a polyhistidine fusion protein in bacteria, bacterial lysates or cells and tissues transfected with a polyHistidinetagged fusion protein expressing vectors.

  • Clonality

    Monoclonal

  • Clone number

    HIS-1

  • Isotype

    IgG2a

  • Research areas

Associated products

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab49746 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 (1)
1/2000. Predicted molecular weight: 7 kDa.
Dot blot
Use at an assay dependent dilution.
ELISA
Use at an assay dependent dilution.
Notes
WB
1/2000. Predicted molecular weight: 7 kDa.
Dot blot
Use at an assay dependent dilution.
ELISA
Use at an assay dependent dilution.

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

References (3)

Publishing research using ab49746? Please let us know so that we can cite the reference in this datasheet.

ab49746 has been referenced in 3 publications.

  • Bottermann M  et al. Complement C4 Prevents Viral Infection through Capsid Inactivation. Cell Host Microbe 25:617-629.e7 (2019). PubMed: 30926239
  • Henche AL  et al. Dissection of key determinants of cleavage activity in signal peptidase III (SPaseIII) PibD. Extremophiles 18:905-13 (2014). PubMed: 25102813
  • Hussack G  et al. Neutralization of clostridium difficile toxin A with single-domain antibodies targeting the cell-receptor binding domain. J Biol Chem : (2011). WB . PubMed: 21216961

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

Inquiry: We are looking into AP conjugated antibodies to detect His-tagged proteins on PIP strips (Echelon). The proteins we use carry the pQE30 Histag (RGS-6xHis). Your anti-6xHis is described to detect tags from pET, pRSET, pTRC. Do you have any information on how it would work with the pQE30 tags? Is it suitable for PIP strips? We plan to use BCIP/NBT detection. What is the detection limit/sensitivity of your AP conjugated antibody with BCIP/NBT? We have not worked with AP cojugated Abs and BCIP/NBT before and therefore appreciate any technical advise you can give us.

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Answer

The 6x His-tag should essentially be the same no matter which vector it is expressed by. The affinity of the antibody is more likely to be affected by the proximity of the His-tag to the recombinant protein (N- or C-terminal or within the reading frame) but as long as the tag is accessible I can see no reason why the antibody would not work.

PIP strips are like a modified dot-blot. This antibody is described as being suitable for dot blots on our website so theoretically it should be suitable for this application.

We have not tested the sensitivity limit of ab49746 with BCIP/NBT but for immunoblotting a minimum working dilution of 1:2,000 is determined using bacteria lysates expressing a recombinant histidine-tagged fusion protein and BCIP/NBT substrate. I would recommend determining the optimal working dilutions by titration.

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Western blot abreview for Anti-6X His tag® antibody [HIS-1] (Alkaline Phosphatase)

 Average
Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (neuronal cell)
Loading amount
10 µg
Specification
neuronal cell
Gel Running Conditions
Reduced Denaturing (12% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Dec 02 2009

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