Our Abpromise guarantee covers the use of ab9695 in the following tested applications.

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

Application Abreviews Notes
WB Use a concentration of 0.1 - 0.2 µg/ml. To detect hEGF by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant hEGF is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions. Western blot bands should be around 6.2 kDa.
ELISA Use a concentration of 0.5 µg/ml. To detect hEGF by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant hEGF.
Neutralising Use a concentration of 0.25 - 0.35 µg/ml. To yield one-half maximal inhibition [ND50] of the biological activity of hEGF (2.0 ng/ml), a concentration of 0.25 - 0.35 µg/ml of this antibody is required.
Sandwich ELISA Use a concentration of 0.5 µg/ml.

For sandwich ELISA, use this antibody as Detection at 0.5µg/ml with ab18635 as Capture.

IHC-P Use a concentration of 1 µg/ml.


  • 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
  • Database links
  • Alternative names
    • Beta urogastrone antibody
    • beta-urogastrone antibody
    • EGF antibody
    • EGF_HUMAN antibody
    • Epidermal growth factor antibody
    • HOMG4 antibody
    • OTTHUMP00000219721 antibody
    • OTTHUMP00000219722 antibody
    • Pro epidermal growth factor antibody
    • URG antibody
    • Urogastrone antibody
    see all


  • ab9695 at 1.0 µg/ml staining normal human skin by IHC-P. The antibody was incubated overnight at 4˚C. An HRP-labeled polymer detection system was used with a non-alcohol soluble AEC chromogen and a proteinase K antigen retrieval.
  • Primary ab9695 was used at a dilution of 0.2 ug/ml.

    Lane 1: 250 ng Recombinant human EGF protein

    Lane 2: 125 ng Recombinant human EGF protein

  • Immunohistochemical analysis of murine forepaw vessel, staining EGF with ab9695.

    Tissue was fixed with formaldehyde and blocked with 1% blocking solution for 15 minutes at room temperature; antigen retrieval was by proteinase K. Samples were incubated with primary antibody (4 µg/ml in BSA in TBS) for 1 hour. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

    See Abreview

  • Standard Curve for EGF (Analyte: ab51082) dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [S-177] to EGF (ab18635) at 5ug/ml and Detector Antibody Rabbit polyclonal to EGF (ab9695) at 0.5ug/ml


This product has been referenced in:
  • Zhang B  et al. Metastasis-associated protein 2 promotes the metastasis of non-small cell lung carcinoma by regulating the ERK/AKT and VEGF signaling pathways. Mol Med Rep 17:4899-4908 (2018). Read more (PubMed: 29393472) »
  • Fang C  et al. Berberine ameliorates neonatal necrotizing enterocolitis by activating the phosphoinositide 3-kinase/protein kinase B signaling pathway. Exp Ther Med 15:3530-3536 (2018). Read more (PubMed: 29545879) »

See all 11 Publications for this product

Customer reviews and Q&As

Western blot
Human Recombinant protein (Human recombinant EGF)
Gel Running Conditions
Reduced Denaturing (16% tricine gel MES)
Loading amount
0.002 µg
Human recombinant EGF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Julien Chevalier

Verified customer

Submitted May 26 2015

Western blot
Loading amount
100000 cells
Gel Running Conditions
Reduced Denaturing (4-10%)
Orthopoxvirus Serum (BHK-21 cells)
BHK-21 cells
Infected with Western Reserve Strain of vaccinia virus
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C

Dr. B Ward

Verified customer

Submitted Oct 22 2013

Thank you very much for your inquiry.
The immunogen for ab9695 was the full length recombinant hEGF (human Epidermal Growth Factor) and it therefore impossible to determine an epitope.
It is possible to determine experimentally if the antibod...

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I am sorry for the delay in getting back to you in regards to the antigen retrieval protocol used with the anti-EGF antibody (ab9695).
Please find the protocol attached. Due to the type of tissue used in this testing of ab9695 (human skin tissue), ...

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Thank you for contacting us yesterday and your interest in our products.
As discussed over the phone I have been looking into what may serve as suitable positive controls with the anti-EGF antibody (ab9695). As discussed over the phone, we have use...

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Thank you for your answer.

We guarantee the ab9695 for WB under either reducing or non-reducing conditions. The free of charge replacement will also be covered by our Abcam guarantee so if you would experience problems we would be pleased to h...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Mouse Tissue sections (Forepaw Vessel)
Forepaw Vessel
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Prot K
Blocking step
DAKO PB as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Nov 30 2012

Thank you for contacting us.
I am sorry to hear that the antibodies are not working as expected.
As we discussed over the phone, if you please could try the no primary control, this would help us a lot in determining whether the problem comes...

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Thakn you for your enquiry. I can confirm the following procedure was used to laboratory test this antibody on skin samples to provide the image on the datasheet. Please note this should be a guideline only and may require further optimization in yo...

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Thank you for your enquiry. I have been in touch with the source of this antibody and I can tell you that ab9695 has been tested and shown to neutralise the specific activity of the antigen (EGF). The neutralisation is measured by ND50. ND50 is the...

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