Overview

Properties

  • FormLyophilised:Reconstitute with 200µl of sterile water.
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles.
  • Storage bufferPBS, pH 7.4, no preservative, sterile filtered
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • Isotypeunknown
  • Light chain typeunknown
  • Research Areas

Applications

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. Can be paired for Sandwich ELISA with Mouse monoclonal [S-177] to EGF (ab18635). 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.

Target

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

Anti-EGF antibody images

  • All lanes : Anti-EGF antibody (ab9695) at 0.2 µg/ml

    Lane 1 : Human EGF full length protein (ab55483) at 0.1 µg
    Lane 2 : Human EGF full length protein (ab55483) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 1 minute
  • 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.
  • 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
  • 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

References for Anti-EGF antibody (ab9695)

This product has been referenced in:
  • Kumagai K  et al. Up-regulation of EGF receptor and its ligands, AREG, EREG, and HB-EGF in oral lichen planus. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 110:748-54 (2010). IHC-P ; Human . Read more (PubMed: 20952227) »

See 1 Publication for this product

Product Wall

Abreviews
Application Western blot
Loading amount 100000 cells
Gel Running Conditions Reduced Denaturing (4-10%)
Sample Orthopoxvirus Serum (BHK-21 cells)
Specification BHK-21 cells
Treatment 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
Username

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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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Forepaw Vessel)
Specification Forepaw Vessel
Fixative Formaldehyde
Antigen retrieval step Enzymatic - Buffer/Enzyme Used: Prot K
Permeabilization No
Blocking step DAKO PB as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
Username

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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"