• Nature
  • Source
    Escherichia coli
  • Amino Acid Sequence
    • Accession
    • Species
    • Sequence
    • 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


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.

  • 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.

  • Applications

    Western blot

    Cellular Activation



  • Endotoxin level
    < 0.010 Eu/µg
  • Purity
    >= 98 % SDS-PAGE.
    Sterile filtered. Greater than 98% pure by HPLC analyses.
  • Form
  • 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
    • 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
    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
  • Information by UniProt


  • 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.

    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  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.


This product has been referenced in:
See all 3 Publications for this product

Customer reviews and Q&As

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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 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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Submitted Jun 25 2010

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