Overview

  • Product name
    Anti-EGF antibody [EGF-10]
    See all EGF primary antibodies
  • Description
    Mouse monoclonal [EGF-10] to EGF
  • Host species
    Mouse
  • Tested applications
    Suitable for: Dot blot, IHC-Fr, ELISA, RIA, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse
  • Immunogen

    Recombinant full length protein (Human).

  • General notes
    If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use.

Properties

Applications

Our Abpromise guarantee covers the use of ab10409 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
Dot blot Use a concentration of 0.1 µg/ml.
IHC-Fr 1/500. PubMed: 17475821(see Abreview)
ELISA Use at an assay dependent concentration.
RIA Use at an assay dependent concentration.
IHC-P 1/500.

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

  • ab10409 staining EGF in human normal skin tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with formaldehyde. Samples were blocked with 10% serum for 30 minutes at 21°C followed by incubation with the primary antibody at a 1/500 dilution for 30 minutes at 21°C. An undiluted HRP-conjugated goat polyclonal was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Lindenmeyer MT  et al. Interstitial vascular rarefaction and reduced VEGF-A expression in human diabetic nephropathy. J Am Soc Nephrol 18:1765-76 (2007). IHC-P, IHC-Fr ; Human . Read more (PubMed: 17475821) »
See 1 Publication for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ***** for ab9695. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

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Question
Answer

Thank you for contacting Abcam regarding ab10409. I have confirmed the following protocol was used in IHC-P: Heat Induced Epitope Retrieval (HIER) is performed by heating the slides immersed in retrieval solution: 10 mM sodium citrate buffer, pH 6.0, with 1 mM EDTA, at 125 °C for 4 minutes in a pressure boiler. After boiling is completed, slides remain in the pressure boiler and are allowed to cool down to 90 °C. The total processing time is ~45 minutes. Staining was performed using a Lab Vision Autostainer 480.  All incubations were performed at RT and all reagents are applied at a volume of 300ul. 1.  Rinse in wash buffer - PBS containing 0.2% Tween 20 2.  Incubate with Ultra V Block 5 minutes 3.  Rinse 2x in wash buffer 4.  Incubate with primary antibody diluted 1:500 for 30 minutes 5.  Rinse 3X in wash buffer 6.  Incubate with HRP labeled polymer for 30 minutes 7.  Rinse 2X in wash buffer 8.  Develop using DAB 9.  Rinse 2X in distilled water 10.  Counterstain I hope this information is helpful.  Please do not hesitate to contact me if you have any additional questions.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Normal Skin)
Specification
Normal Skin
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jan 11 2011

Answer

Thank you for your enquiry. We tested ab10409 with a recombinant EGF sample, which is available from Sigma I believe, catalogue number E9644. Please do not hesitate to contact me again if you require further information,

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Answer

Thanks for your enquiry regarding ab10409. Unfortunately we do not currently offer free trial size batches of antibodies to test. I should take this opportunity to mention that this antibody may not recognise rat EGF since the antibody is known not to recognise mouse EGF. Good luck with your research!

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Answer

Thank you for your note. We are very sorry to hear that you are still struggling with this antibody and even the replacement vial does not perform as it should do. We can offer you either a credit note or a full refund; please do let us know how to proceed. We would be grateful if you could confirm your Purchase Order Number or Abcam Order Number. We apologize for any inconvenience caused and we look forward to hearing from you soon.

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Question

ANTIBODY CODE ab10409 BATCH NUMBER 62221 DESCRIPTION OF THE PROBLEM No signal at all SAMPLE human blood, plasma and brain. Whole protein extraction. PRIMARY ANTIBODY abcam. mouse anti EGF (monoclonal). 1-5 ug/ml in TBST and 5% BSA & 1% goat serum. Incubation at 4?C overnight. SECONDARY ANTIBODY Goat anti mouse (Pierce). 1:500 in 5%non-fat milk & 1% goat serum in TBST DETECTION METHOD SuperSignal West Dura, Pierce. POSITIVE AND NEGATIVE CONTROLS USED No ANTIBODY STORAGE CONDITIONS alicuoted and stored at -20?C SAMPLE PREPARATION Extraction Buffer: Tris-HCl 20mM, Sucrose 10%, EDTA 1mM, EGTA 1mM & inhibitors complete mini EDTA-free tablets (Roche) Also tryed NP40 with inhibitors as extraction buffer. Proteins were mixed with sample buffer 4x:Tris 250mM, glycerol 20%, b-mercaptoethanol 20%, bromophenol blue 0.002% and SDS 8%, and then they were heated at 100?C for 5 min, before loaded. AMOUNT OF PROTEIN LOADED 10-50 ug ELECTROPHORESIS/GEL CONDITIONS SDS poliacrilamide gel electrophoresis. 12% TRANSFER AND BLOCKING CONDITIONS semi-dry transference, 15min per minigel 15V. Non-fat milk as blocking agent 1h at room T?. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Diffent samples and different protein concentrations. Variation in primary antibody concentration and I have increased incubation time up to 24 h. ADDITIONAL NOTES I have never obtained a signal. I have had no problems with other primary antibodies developed mouse, using the same method. I have chequed the transference with Ponceau Red, and it was always OK. Also loaded prestained protein standards.

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Answer

Thank you for your enquiry and forwarding your detailed protocol to us. We are very sorry to hear that you are having problem with this antibody. We would suggest loading purified human EGF (1-100 ng) as positive control along with the samples. Should you still have problem with this antibody, then please do not hesitate to contact us again. We can offer you one replacement vial of ab10409 - free of charge. Looking at your order details and shipping record on FedEx website, it took 4 days for you to receive this item. It may well be that the antibody has gone off during long shipping. Please do let us know if you prefer to get a new vial. Best regards

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Answer

We are not aware of any publications where this antibody is used. However, if you have a look at the reviews section of the online datasheet, you'll see that another Abcam customer used this antibody in IHC on formalin-fixed paraffin-embedded human autopsy tissue and recommended a 1:500 dilution. Furthermore, they have included their protocol. You do not give any details of the conditions of your experiments, but perhaps the concentration at which you are using the anitbody is too high?

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