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Bioactive grade

Recombinant human EGF protein (Animal Free) (ab9697)

Reviews (1)Q&A (1)References (9)
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

Achieve higher consistency and quality standards with a premium grade bioactive protein

Product image
Recombinant human EGF protein (Active) (ab259398)
  • High batch-to-batch consistency
  • Optimal bioactivity
  • Guaranteed identical to human native proteins
  • >95% purity
  • Ultra-low endotoxin levels: <0.005 Eu/µg
  • Carrier and tag free

Description

  • Product name

    Recombinant human EGF protein (Animal Free)
    See all EGF proteins and peptides

  • Biological activity

    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.

  • Purity

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

  • Endotoxin level

    < 0.010 Eu/µg

  • Expression system

    Escherichia coli

  • Accession

    Q6QBS2

  • Protein length

    Full length protein

  • Animal free

    Yes

  • Nature

    Recombinant

    • Species

      Human

    • Sequence

      NSDSECPLSH DGYCLHDGVC MYIEALDKYA CNCVVGYIGE RCQYRDLKWW ELR

    • Predicted molecular weight

      6 kDa

    • Amino acids

      971 to 1023

    • Additional sequence information

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

Associated products

Specifications

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.

  • Applications

    Western blot

    Cellular Activation

    SDS-PAGE

    HPLC

  • Form

    Lyophilized

  • Additional notes

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

Preparation and Storage

  • Stability and Storage

    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.

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

General Info

  • Alternative names

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

  • Function

    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).

  • Tissue specificity

    Expressed in kidney, salivary gland, cerebrum and prostate.

  • Involvement in disease

    Hypomagnesemia 4

  • Sequence similarities

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

  • Post-translational
    modifications

    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).

  • Cellular localization

    Membrane.
  • Information by UniProt

Images

  • 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.

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

References (9)

Publishing research using ab9697? Please let us know so that we can cite the reference in this datasheet.

ab9697 has been referenced in 9 publications.

  • Cao X  et al. Alt-RPL36 downregulates the PI3K-AKT-mTOR signaling pathway by interacting with TMEM24. Nat Commun 12:508 (2021). PubMed: 33479206
  • Cui N  et al. Hexokinase 2 Promotes Cell Growth and Tumor Formation Through the Raf/MEK/ERK Signaling Pathway in Cervical Cancer. Front Oncol 10:581208 (2020). PubMed: 33324557
  • 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

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 25 2010

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.

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