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    e-coli-positive-control-escherichia-coli--whole-cell-lysate-expressing-6x-his-tag-protein-ab2431.pdf

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Microbiology Organism Bacteria Gram negative Bacteria Escherichia coli
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E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431)

  • Datasheet
  • SDS
Submit a review Q&A (6)References (1)

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Western blot - E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431)

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    Overview

    • Product name

      E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein
    • General notes


      This lysate can be used as a positive control for Western blots using lysates containing 6X-His tagged proteins.

    • Tested applications

      Suitable for: WBmore details

    Properties

    • Mycoplasma free

      Yes
    • Form

      Lyophilized:After the lyophilised pellet is dissolved in 250 ul SDS-gel sample buffer (1 mg/ml protein lysate) it can be used direct for sample loading and then stored at –20°C. The sample can be reused several times
    • Storage instructions

      Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
    • Concentration information loading...
    • Lysate notes

      The cell lysate was prepared from E.coli (Tuner [De3]) expressing a 14.2 kDa protein containing a C-terminal 6xHis tag.

    • Research areas

      • Microbiology
      • Organism
      • Bacteria
      • Gram negative Bacteria
      • Escherichia coli
      • Kits/ Lysates/ Other
      • Lysates
      • Whole Cell Lysates
      • Arabidopsis / Miscellaneous

    Associated products

      Applications

      Our Abpromise guarantee covers the use of ab2431 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 Use at an assay dependent dilution. This E. coli (strain BL21)lysate contains a recombinant C-terminal 6x-His tagged protein of 17 kDa. The his-tagged protein is monomeric and will be detected under reducing and non-reducing conditions. Use 5-20 µg per lane of a mini-gel at 12-15 % SDS-PAGE. After the lyophilised pellet is dissolved in 250 ul SDS-gel sample buffer (1 mg/ml protein lysate) it can be used direct for sample loading and than stored at -20°C. The sample can be reused several times.

      Images

      • Western blot - E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431)
        Western blot - E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431)
        All lanes : Rabbit anti-His-tag antibody

        Lane 1 : E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431) at 2 µg
        Lane 2 : E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431) at 5 µg
        Lane 3 : E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431) at 10 µg
        Lane 4 : E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431) at 15 µg
        Lane 5 : E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431) at 20 µg

        Secondary
        All lanes : Goat anti-rabbit antibody (AP-conjugated)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
      • SDS
    • References (1)

      Publishing research using ab2431? Please let us know so that we can cite the reference in this datasheet.

      ab2431 has been referenced in 1 publication.

      • Wright JC  et al. Enhanced peptide identification by electron transfer dissociation using an improved mascot percolator. Mol Cell Proteomics : (2012). PubMed: 22493177

      Customer reviews and Q&As

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

      1-6 of 6 Abreviews or Q&A

      Question

      What is the protocol and lysis buffer used to produce this e coli lysate?

      Read More

      Abcam community

      Verified customer

      Asked on Nov 27 2012

      Answer

      E coli was grown in LB-Amp medium.

      Read More

      Abcam Scientific Support

      Answered on Nov 27 2012

      Question

      What is the protocol and lysis buffer used to produce this e coli lysate?

      Read More

      Abcam community

      Verified customer

      Asked on Nov 19 2012

      Answer

      The E. coli was cultured using a standard protocol and harvested by centrifugation. The pellet was resuspended in 50mM Tris, 10mM EDTA, pH8.0, treated with ultrasonic sound, centrifuged and the supernatant was finally filtered (0.22µm).

      Read More

      Abcam Scientific Support

      Answered on Nov 19 2012

      Question

      Are these total cell lysates and how they were lysed

      Read More

      Abcam community

      Verified customer

      Asked on Mar 20 2012

      Answer

      Je peux vous confirmer que les cellules E. coli du produit ab2431 ont été préparé dans un tampon contenant 50mM Tris, 10mM EDTA, pH 8.0 et lysé par sonification ensuite. Le lysat était ensuite aliquoté et lyophilisé.

      Read More

      Abcam Scientific Support

      Answered on Mar 20 2012

      Question

      What is the recipe for the SDS-gel sample buffer that you recommend reconstituting the lyophilized product in? What is the concentration, 1x or 4x? Also, I'm using this as a positive control in WB, not SDS-PAGE. Should I reconstitute in something else?

      Read More

      Abcam community

      Verified customer

      Asked on Jun 18 2010

      Answer

      Thank you for your inquiry. We use the following 4X sample buffer: 4 mL mQ H2O 1 mL 0.5M Tris-Cl, pH 6.8 0.8 mL Glycerol 1.6 mL 10% SDS (w/v) 0.2 mL Bromphenol blue + 400µl Mercaptoethanol (for reduced gels) If you reconstitute the lysate directly in sample buffer we would recommend to use 1X. The total protein lysate is suggested to use as a standard or positive control when expressing 6H-tagged proteins in E. coli. When analyzing the total protein lysate on a gel by Western blotting a prominent band of 17 kDa will interact with anti-his-tag antibodies. The His-tagged protein is monomeric and will be detected under reducing and non-reducing conditions. It is recommended to use 5 mg – 20 mg protein lysate per lane for SDS-PAGE. After the lyophilized pellet is dissolved in 250 µl SDS-gel sample buffer (1 mg/ml protein lysate) it can be used direct for sample loading and then stored at –20°C. The sample can be reused several times..

      Read More

      Abcam Scientific Support

      Answered on Jun 18 2010

      Question

      Customer is interested in this lysate (ab2431) but is concerned about the other bands shown on the gel, would like more information. Are they non-specific, etc?

      Read More

      Abcam community

      Verified customer

      Asked on Feb 05 2004

      Answer

      The E. coli lysate containing the his-tag protein is a whole bacterial cell pellet lysate. The amount of bands detected with a his-tag antibody depends on the quality of the his-tag antibody and is therefore antibody dependent.

      Read More

      Abcam Scientific Support

      Answered on Feb 06 2004

      Question

      I am looking for a positive control on a slot blot assay. Will your his tag E.coli lysate work in a slot blot? Thanks.

      Read More

      Abcam community

      Verified customer

      Asked on Oct 13 2003

      Answer

      To our knowledge, this antibody has yet to be tested in this application. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.

      Read More

      Asdf Edo

      Abcam Scientific Support

      Answered on Oct 13 2003

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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