Overview

Properties

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.

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

  • 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
  • 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

Images

  • 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

References

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

1-10 of 10 Abreviews or Q&A

Abreviews
Application
Western blot
Sample
Human Recombinant protein (Human recombinant EGF)
Gel Running Conditions
Reduced Denaturing (16% tricine gel MES)
Loading amount
0.002 µg
Specification
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

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

Dr. B Ward

Verified customer

Submitted Oct 22 2013

Answer

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 antibody blocks the binding site of EGFR, but this has not been done and also would have to be repeated with every new immunization round (for every batch of antibody).
To our knowledge, ab9695 has not been tested in immunoprecipitation and therefore is also not covered by our guarantee for this application. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.
Simply follow these easy steps below to apply for our AbTrial Program:
1. Reply to this email, letting us know you are interested in testing this product.
2. Our scientists will email you an inactive personal discount code for the value of the product.
3. Purchase and test the product at the regular price.
4. Submit your results, including your discount code in the additional notes section of your Abreview.
5. Once the Abreview is submitted, the discount code will become active.
6. Apply your discount code on your next order to receive that value off.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

Read More

Answer

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), the HEIR would not have been suitable as it can causes the skin to lift off of the slide. This does not mean that it will not be suitable with other tissue samples.
I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

Read More

Answer

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 used the antibody to stain normal human skin as well as murine forepaw vessel. The protein seems to also be expressed in kidney, salivary gland, cerebrum and prostate, so if you have access to any of these tissues they could serve as a control.
If I can be of any further assistance, please do not hesitate to let me know.
Until then, I wish you all the best with your research.

Read More

Answer

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 help in this case as well.

Please let me know about your decision in this case. i look forward hearing from you if you decide to receive the ab9695 as a replacement.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Nov 30 2012

Answer

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 from the secondary antibody alone, or if there are issues with the primary antibody and isotype control as well.
Please let me know your results.
I look forward to hear back from you and assist you further in resolving his problem.

Read More

Answer

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 your laboratory: Antibody concentration: 1.0 μg/ml Tissue: Normal Human Skin (formalin fixed, paraffin embedded) Protocol: Rabbit HRP Polymer Kit (AEC chromogen) Antigen Retrieval: Proteinase K Incubation Times: 10 min Proteinase K 20 min 1X PBS/ 0.2% Trinton X 100 30 min Fc Receptor Block 10 min Sniper Protein Block Overnight Primary Antibody @ 4°C 10 min Peroxidase Block 20 min Rabbit Probe 20 min Rabbit Polymer HRP 5-10 min Romulin AEC chromogen Results: This antibody stained normal skin with 4+ apocrine gland epithelial and secretory cells. Everything else is negative. Ideal staining is at 1.0 μg/ml, yet this antibody concentration range was tested from 1.0 μg/ml- 0.25 μg/ml. I can confirm that any publications that we are currently aware of will be listed on the datasheet. I'm sorry there are none to provide on this occasion for ab9695 EGF antibody. In many cases, regrettably we do not have access to articles mentioning our products, or subscriptions to the journals that publish them. I hope this will be helpful. Should you have any further questions, please do not hesitate to contact us.

Read More

Answer

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 amount of antibody to neutralize 50% of the biological activity of the specific concentration of antigen. Unfortunately this antibody is not supplied conjugated to flurophores. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

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

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