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

Submit a review Q&A (6)References (3)
Western blot - E. coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 6X His tag protein (ab2431)

    製品の概要

    • 製品名

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

    • 特記事項


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

    • アプリケーション

      適用あり: WBmore details

    製品の特性

    • Mycoplasma free

      Yes

    • 製品の状態

      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

    • 保存方法

      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...

    • ライゼート 備考

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

    • 研究分野

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab2431の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    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.
    特記事項
    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.

    画像

    • 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)

    プロトコール

    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

    参考文献 (3)

    ab2431 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab2431 は 3 報の論文で使用されています。

    • Miao K  et al. NOTCH1 activation compensates BRCA1 deficiency and promotes triple-negative breast cancer formation. Nat Commun 11:3256 (2020). PubMed: 32591500
    • Imai K  et al. Biological Effects of Shikonin in Human Gingival Fibroblasts via ERK 1/2 Signaling Pathway. Molecules 24:N/A (2019). PubMed: 31574951
    • Wright JC  et al. Enhanced peptide identification by electron transfer dissociation using an improved mascot percolator. Mol Cell Proteomics : (2012). PubMed: 22493177

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    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
    Answer

    E coli was grown in LB-Amp medium.

    Read More

    Question

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

    Read More
    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

    Question

    Are these total cell lysates and how they were lysed

    Read More
    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

    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
    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

    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
    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

    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
    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

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