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    products/primary-antibodies/egf-antibody-ab9695.pdf

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Signal Transduction Growth Factors/Hormones EGF
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Anti-EGF antibody (ab9695)

  • Datasheet
  • SDS
Reviews (3)Q&A (7)References (28)

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Abpromise

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Find out more.

Western blot - Anti-EGF antibody (ab9695)
  • Sandwich ELISA - Anti-EGF antibody (ab9695)

Key features and details

  • Rabbit polyclonal to EGF
  • Suitable for: WB, IHC-P, ELISA, Neutralising, Sandwich ELISA
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-EGF antibody [EPR19899] (ab206423)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-EGF antibody
    See all EGF primary antibodies
  • Description

    Rabbit polyclonal to EGF
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ELISA, Neutralising, Sandwich ELISAmore details
  • Species reactivity

    Reacts with: Human, Recombinant fragment
  • Immunogen

    Recombinant full length protein corresponding to Human EGF. Highly pure (>98%) recombinant hEGF (human Epidermal Growth Factor)
    Database link: P01133

  • Positive control

    • Purchase matching WB positive control:Recombinant human EGF protein (Animal Free)
  • General notes

    Although some customers have been successful in IHC we no longer batch test in this application. For IHC validate antibody please see ab184265.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Lyophilized:Reconstitute with 200µl of sterile water.
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer

    No preservative, sterile filtered
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Light chain type

    unknown
  • Research areas

    • Signal Transduction
    • Growth Factors/Hormones
    • EGF
    • Cell Biology
    • Cell Cycle
    • Cell differentiation
    • Cancer
    • Growth factors
    • EGF
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Growth factors and hormones ELISA kits
    • Neuroscience
    • Processes

Associated products

  • Positive Controls

    • Recombinant human EGF protein (Animal Free) (ab9697)
  • Recombinant Protein

    • Recombinant human EGF protein (Active) (ab259398)

Applications

The Abpromise guarantee

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 (2)
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.
IHC-P (1)
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.

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 at an assay dependent concentration.

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 - 2 µg/ml.

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

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.
IHC-P
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.

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 at an assay dependent concentration.

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 - 2 µg/ml.

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

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.
  • Target information above from: UniProt accession P01133 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1950 Human
    • Omim: 131530 Human
    • SwissProt: P01133 Human
    • Unigene: 419815 Human
    • 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

    • Western blot - Anti-EGF antibody (ab9695)
      Western blot - Anti-EGF antibody (ab9695)

      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

    • Sandwich ELISA - Anti-EGF antibody (ab9695)
      Sandwich ELISA - Anti-EGF antibody (ab9695)

      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

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (28)

    Publishing research using ab9695? Please let us know so that we can cite the reference in this datasheet.

    ab9695 has been referenced in 28 publications.

    • Ma M  et al. SMAD-specific E3 ubiquitin ligase 2 promotes angiogenesis by facilitating PTX3 degradation in MSCs from patients with ankylosing spondylitis. Stem Cells 39:581-599 (2021). PubMed: 33547700
    • Wu M  et al. P25 Gene Knockout Contributes to Human Epidermal Growth Factor Production in Transgenic Silkworms. Int J Mol Sci 22:N/A (2021). PubMed: 33800168
    • Mao Y  et al. The Overexpression of Epidermal Growth Factor (EGF) in HaCaT Cells Promotes the Proliferation, Migration, Invasion and Transdifferentiation to Epidermal Stem Cell Immunophenotyping of Adipose-Derived Stem Cells (ADSCs). Int J Stem Cells 13:93-103 (2020). PubMed: 32114740
    • Praktiknjo SD  et al. Tracing tumorigenesis in a solid tumor model at single-cell resolution. Nat Commun 11:991 (2020). PubMed: 32080185
    • Haque ASMR  et al. CD206+ tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production. Sci Rep 9:14611 (2019). PubMed: 31601953
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-EGF antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Julien Chevalier

    Verified customer

    Submitted May 26 2015

    Western blot abreview for Anti-EGF antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. B Ward

    Verified customer

    Submitted Oct 22 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EGF antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 30 2012

    Question

    Dear Sirs



    We have purchased anti-EGF antibody (ab9695)in your company. I would like to kindly ask if you could provide more information regarding epitope of this antibody? We would like to know if the anti-EGF ab is suitable for co-immunoprecipitaion with EGFR? Does the antibody binding site co-localize with EGFR binding site, thus blocking interaction between EGF and its receptor EGFR?



    Thank you for your help



    Best regards

    Read More

    Abcam community

    Verified customer

    Asked on Feb 11 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

    Abcam Scientific Support

    Answered on Feb 11 2013

    Question

    Thank you for your help.
    We have managed to get signal on initial trial IHC. However, only a very few cells are staining.
    This may be due to limited expression in our tissue, but we wish to try to optimise to see if signal can be improved.
    Your data sheet recommends proteinase K antigen retrieval - we used citrate buffer for the initial trial as per your general IHC notes on the Abcam website. Can you please provide exact recommended conditions for the proteinase K method?
    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 08 2013

    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

    Abcam Scientific Support

    Answered on Jan 08 2013

    Question

    Phone call requesting suitable positive control when using the anti-EGF antibody (ab9695) in IHC-P

    Read More

    Abcam community

    Verified customer

    Asked on Dec 18 2012

    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

    Abcam Scientific Support

    Answered on Dec 18 2012

    Question

    Thank you, yes, this email is a lot easier to understand! Thank you for the offer of a replacement antibody. After looking through the list, ab9695 looks like it would suit me well. Could you please confirm for me whether or not it has been tested under denaturing conditions, or if it will only detect the non-denatured protein?
    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 13 2012

    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 Scientific Support

    Answered on Dec 13 2012

    Question

    IHC-P
    mouse tumor tissue (from lung) with human cell line injected, some muscle tissue attached as well
    sees high background for human cellswith priamry Ab as well as isotype ctrl, but also for muscle (does not contain any EGF)
    Ab: 1 ug/ml 3h RT
    block: horse serum + chicken egg white albumin
    AR: none + proteinase K
    2nd (ab96879)

    suggested: try no primary crtl to determine if problem is secondary

    Read More

    Abcam community

    Verified customer

    Asked on May 15 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

    Abcam Scientific Support

    Answered on May 15 2012

    Question

    Hi, I recently purchased EGF antibody (ab9695). I am trying to find the specific referrence in PUBMED of how EGF was used to stain skin with immunohistochemistry. is it possible to sent me that rererence or the papaer so that I can read their staining methods? Thanks so much in advance for looking into it! Ioanna

    Read More

    Abcam community

    Verified customer

    Asked on Sep 28 2010

    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

    Abcam Scientific Support

    Answered on Sep 28 2010

    Question

    Can you tell me what "neutralisation" means. Does this mean that it prevents the binding effects of the ligand, replaces the effects of the ligand or another biological effect. Also can you tell me whether this antibody comes conjugated to a flurophore?

    Read More

    Abcam community

    Verified customer

    Asked on May 11 2006

    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

    Abcam Scientific Support

    Answered on May 12 2006

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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