Active human EGF full length protein (ab9697)

Overview

Description

  • NatureRecombinant
  • SourceE. coli
  • Amino Acid Sequence
    • SpeciesHuman
    • SequenceRecombinant Human EGF is a 6.2 kDa protein containing 53 amino acid residues: NSDSECPLSH DGYCLHDGVC MYIEALDKYA CNCVVGYIGE RCQYRDLKWW ELR
    • Amino acids0 to 0

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

  • FormLyophilised
  • Additional NotesManufactured using all non-animal reagents.
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    n/a

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

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

General info

  • Alternative names
    • Beta urogastrone
    • EGF
    • Egf
    • EGF_HUMAN
    • Epidermal Growth Factor
    • Epidermal Growth Factor
    • HOMG4
    • OTTHUMP00000219721
    • OTTHUMP00000219722
    • Pro epidermal growth factor
    • URG
    • Urogastrone
    see all
  • FunctionEGF 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.
  • Tissue specificityExpressed in kidney, salivary gland, cerebrum and prostate.
  • Involvement in diseaseDefects in EGF are the cause of hypomagnesemia type 4 (HOMG4) [MIM:611718]; also known as renal hypomagnesemia normocalciuric. HOMG4 is a disorder characterized by massive renal hypomagnesemia and normal levels of serum calcium and calcium excretion. Clinical features include seizures, mild-to mederate psychomotor retardation, and brisk tendon reflexes.
  • Sequence similaritiesContains 9 EGF-like domains.
    Contains 9 LDL-receptor class B repeats.
  • Cellular localizationMembrane.
  • Information by UniProt

Active human EGF full length protein images

  • Lane 1 :
    Lane 2 :
    Lane 3 :

    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
    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

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Active human EGF full length protein (ab9697)

ab9697 has not yet been referenced specifically in any publications.

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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 powde...

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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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Submitted Jun 25 2010

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