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

Q&A (6)Specific References (1)

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

  • Form
    Lyophilised: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

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

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

    Western blot of 9 µg of E. coli BL21 lysate containing a 17kDa 6x-His tagged protein.  Mouse monoclonal to His tag (4D11 - ab5000) is used at 1:1000.

References

This product has been referenced in:
  • Wright JC  et al. Enhanced peptide identification by electron transfer dissociation using an improved mascot percolator. Mol Cell Proteomics : (2012). Read more (PubMed: 22493177) »

See 1 Publication for this product

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

Customer reviews and Q&As

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6 Q&A

10
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What is the protocol and lysis buffer used to produce this e coli lysate?

E coli was grown in LB-Amp medium.

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

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).

Are these total cell lysates and how they were lysed

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.

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 ...

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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 reconstitu...

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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?

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.

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.

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 fo...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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