Bioactive grade

Recombinant human EGF protein (Animal Free) (ab9697)

Reviews (1)Q&A (1)References (7)
Western blot - Recombinant human EGF protein (Animal Free) (ab9697)
  • Immunocytochemistry/ Immunofluorescence - Recombinant human EGF protein (Animal Free) (ab9697)

Key features and details

  • Expression system: Escherichia coli
  • Purity: >= 98% SDS-PAGE
  • Endotoxin level: < 0.010 Eu/µg
  • Active: Yes
  • Suitable for: WB, Cellular Activation, SDS-PAGE, HPLC

製品の詳細

  • 製品名

    Recombinant human EGF protein (Animal Free)
    EGF タンパク質・ペプチド 製品一覧

  • 生理活性

    The ED50,as determined by a cell proliferation assay using balb/c 3T3 cells, is ≤ 0.1 ng/ml, corresponding to a specific activity of ≥ 1 x 107 units/mg.

  • 精製度

    >= 98 % SDS-PAGE.
    Sterile filtered. Greater than 98% pure by HPLC analyses.

  • エンドトキシン・レベル

    < 0.010 Eu/µg

  • 発現系

    Escherichia coli

  • アクセッション番号

    Q6QBS2

  • タンパク質長

    Full length protein

  • Animal free

    Yes

  • 由来

    Recombinant

    • 生物種

      Human

    • 配列

      NSDSECPLSH DGYCLHDGVC MYIEALDKYA CNCVVGYIGE RCQYRDLKWW ELR

    • 予測される分子量

      6 kDa

    • 領域

      971 to 1023

    • 配列の追加情報

      ab9697 is a globular protein containing 53 amino acid residues, including 2 intramolecular disulfide bonds.

関連製品

特性

Our Abpromise guarantee covers the use of ab9697 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • アプリケーション

    Western blot

    Cellular Activation

    SDS-PAGE

    HPLC

  • 製品の状態

    Lyophilized

  • 備考

    Manufactured using all non-animal reagents.
  • Concentration information loading...

前処理および保存

  • 保存方法および安定性

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • 再構成
    Centrifuge the vial prior to opening. Reconstitute in 500ul sterile filtered water to a concentration of 1mg/ml.

関連情報

  • 別名

    • Beta urogastrone
    • beta-urogastrone
    • EGF
    • EGF_HUMAN
    • Epidermal growth factor
    • HOMG4
    • OTTHUMP00000219721
    • OTTHUMP00000219722
    • Pro epidermal growth factor
    • URG
    • Urogastrone
    see all

  • 機能

    EGF stimulates the growth of various epidermal and epithelial tissues in vivo and in vitro and of some fibroblasts in cell culture. Magnesiotropic hormone that stimulates magnesium reabsorption in the renal distal convoluted tubule via engagement of EGFR and activation of the magnesium channel TRPM6. Can induce neurite outgrowth in motoneurons of the pond snail Lymnaea stagnalis in vitro (PubMed:10964941).

  • 組織特異性

    Expressed in kidney, salivary gland, cerebrum and prostate.

  • 関連疾患

    Hypomagnesemia 4

  • 配列類似性

    Contains 9 EGF-like domains.
    Contains 9 LDL-receptor class B repeats.

  • 翻訳後修飾

    O-glycosylated with core 1-like and core 2-like glycans. It is uncertain if Ser-954 or Thr-955 is O-glycosylated. The modification here shows glycan heterogeneity: HexHexNAc (major) and Hex2HexNAc2 (minor).

  • 細胞内局在

    Membrane.
  • Information by UniProt

画像

  • Western blot - Recombinant human EGF protein (Animal Free) (ab9697)
    Western blot - Recombinant human EGF protein (Animal Free) (ab9697)This image is courtesy of an anonymous Abreview
    All lanes :

    Lane 1 : Whole cell lysate of human skin fibroblasts starved overnight in serum-free medium
    Lane 2 : Whole cell lysate of human skin fibroblasts starved overnight in serum-free medium and then incubated for 30 min with 25 ng/ml active EGF
    Lane 3 : Whole cell lysate of human skin fibroblasts starved overnight in serum-free medium and then incubated for 30 min with 50 ng/ml active EGF

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : An HRP-conjugatede Goat anti-rabbit IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Observed band size: 42,44 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking Step: 5% milk for 1 hour at 25°C

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Recombinant human EGF protein (Animal Free) (ab9697)
    Immunocytochemistry/ Immunofluorescence - Recombinant human EGF protein (Animal Free) (ab9697)

    Immunocytochemistry/ Immunofluorescence  analysis of A431 (Human epidermoid carcinoma cell line) cells labeling EGFR with ab9697 at 1/100, 3 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, a AlexaFluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000, 2 μg/ml. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclear stain was DAPI (blue).

    The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.

    For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.

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 (7)

ab9697 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。

ab9697 は 7 報の論文で使用されています。

  • Deng QF  et al. Cyclooxygenase-2 mediates gefitinib resistance in non-small cell lung cancer through the EGFR/PI3K/AKT axis. J Cancer 11:3667-3674 (2020). PubMed: 32284763
  • Zheng H  et al. Tropomodulin 3 modulates EGFR-PI3K-AKT signaling to drive hepatocellular carcinoma metastasis. Mol Carcinog 58:1897-1907 (2019). PubMed: 31313392
  • Yuan H  et al. Long noncoding RNA LINC01089 predicts clinical prognosis and inhibits cell proliferation and invasion through the Wnt/ß-catenin signaling pathway in breast cancer. Onco Targets Ther 12:4883-4895 (2019). PubMed: 31417284
  • Elliyanti A  et al. Epidermal Growth Factor and Adenosine Triphosphate Induce Natrium Iodide Symporter Expression in Breast Cancer Cell Lines. Open Access Maced J Med Sci 7:2088-2092 (2019). PubMed: 31456831
  • Jong WS  et al. Application of an E. coli signal sequence as a versatile inclusion body tag. Microb Cell Fact 16:50 (2017). PubMed: 28320377
  • Serrano MJ  et al. EMT and EGFR in CTCs cytokeratin negative non-metastatic breast cancer. Oncotarget 5:7486-97 (2014). PubMed: 25277187
  • Prasse AA  et al. Improvement of an antibody-enzyme coupling yield by enzyme surface supercharging. BMC Biotechnol 14:88 (2014). PubMed: 25326050

レビューと Q&A

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1-2 of 2 Abreviews or Q&A

Question


I found three EGF proteins ab9697 (active), ab91371 (active) and ab51082. What’s the difference between active one and not? And what's the difference between two active ones? Which one may I use it to induce the phosphorylation of EGFR in cells?

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Answer

Thank you for contacting us.
The two active proteins are essentially the same, and both can be used to induce phosphorylation as they have been tested in functional studies. The only difference would be that ab9697 is supplied as lyophilized powder and then you can reconstitute it in water, the other active protein ab91371 is in buffer with 50% Glycerol, PBS, 150mM Sodium chloride, 10mM Sodium phosphate, pH 7.2

Ab51082 has not been tested for biological activity, and is intended to use in SDS-PAGE only.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Western blot Abreview for EGF protein (Active)

 Excellent
Abreviews
abreview image
Application
Western blot

Review text: Human skin fibroblasts (HFF-1) were starved in serum-free medium overnight, and then were incubated 30 min in the presence of either 25 or 50 ng/ml active EGF protein, doses widely used in stem cell culture medium, to test whether EGFR downstream signaling pathway is able to be activated.
Sample: Human Cell lysate - whole cell (skin fibroblasts)
Loading amount: 20 µg
Specification: skin fibroblasts
Treatment: 25 and 50 ng/ml EGF for 30 min
Gel Running Conditions: Non-reduced Denaturing (10%)
Blocking step: Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Other product details
Dilution: 1/1000
Incubation time: 16 hour(s) and 0 minute(s) · Temperature: 4°C

Primary antibody (in addition to 'EGF protein (Active)')
Name: Non-Abcam antibody was used: phospho-p44/42 MAPK (Erk1/2)
Host species: Rabbit
Clonality: Polyclonal
Dilution: 1/1000

Secondary antibody
Name: Non-Abcam antibody was used: anti-rabbit 2nd Ab HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution: 1/10000

Detection
Detection method: ECL
Exposure: 1 second(s)
Bands: Specific: 42/44 kDa

Additional data
Additional Notes: Incubation of human fibroblasts with active EGF protein for short time greatly activated EGFR signaling pathway, as the receptor underwent endocytosis and dramatic phosphorylation of p44/42 MAPK.

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投稿 Jun 25 2010

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