Overview

  • Product name

    Human EGF ELISA Kit
    See all EGF kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 1 pg/ml
  • Range

    0.82 pg/ml - 200 pg/ml
  • Recovery

    95 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 95.52 85% - 106%
    Serum 94.52 83% - 104%
    Plasma 93.76 84% - 105%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s EGF Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human EGF in serum, plasma and cell culture supernatants.

    This assay employs an antibody specific for Human EGF coated on a 96-well plate. Standards and samples are pipetted into the wells and EGF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human EGF antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of EGF bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Optimisation may be required with urine samples.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    600X HRP-Streptavidin Concentrate 1 x 200µl
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human EGF (lyophilized) 2 vials
    EGF Microplate (12 strips x 8 wells) 1 unit
    Recombinant Human EGF Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • 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
  • Alternative names

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

Applications

Our Abpromise guarantee covers the use of ab100504 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Standard curve in Diluent A : mean of duplicates (+/-SD)

  • Standard curve in Diluent B : mean of duplicates (+/-SD)

Protocols

References

This product has been referenced in:

  • Kim SW  et al. Amniotic mesenchymal stem cells enhance wound healing in diabetic NOD/SCID mice through high angiogenic and engraftment capabilities. PLoS One 7:e41105 (2012). Human . Read more (PubMed: 22815931) »
See 1 Publication for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

I am sorry to read that the most recent order of the EGF ELISA gave a poor result compared to previous kits. I have arranged for a free replacement from a different lot, order ***.

To check the status of the order please contact our Customer Service team and reference this number.

We have not received other complaints, aside from the complaint from your lab earlier in the year regarding a precipitate in the standards. I am not sure which reagent is defective in this most recent kit, but it is apparently one of the detection reagents, if the older standards that you tested are still intact and undegraded.

Please let us know if you have other questions or concerns.

Read More

Answer

Thank you for confirming these details and for your update. I am pleased to hear the FOCR kit has worked well. This enquiry has been forwarded to me as my colleague #### is currently away from the office.

As requested, I have now issued the second free of charge replacement with the order number ######.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacementis 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.

Read More

Answer

Thank you for confirming that information.

I have now arranged for the kit to be sent out to you. The order number is xxxxx(Purchase order number FOCR xxxxx).

I hope this kit will work as your previous ones have, however, if you have any problems please do let me know. Please take care to note by how much the HRP-streptavidin component needs to be diluted by with this new kit as it is likely to be the 200 uL of 600x.

I look forward to hearing how you get on.

Read More

Answer

Thank you for providing me with that information.

Could you please check how much HRP-streptavidin is in the unused kit which you have? Also, for both of these kits, what dilution did the protocol sent witheach kit advise?

Many thanks for your continued cooperation.

Read More

Question
Answer



We may have identified what the difference may be. Could you confirm, how did you dilute the HRP-streptavidin concentrate provided with the kit?

We have recently modified this reagent and the old kit contained 8 uL of 30,000x HRP-streptavidin (which needed to be diluted by30,000 to obtain the working solution) whilst the newer kits contain 200 uL of 600x (which needs to be diluted600 fold to obtain the working solution). The necessary dilution should be described in the protocol booklet provided with the kit.

Could you please confirm whether you followed the protocol provided with the kit, or the protocol provided with the older kit (purchased in November)?

Many thanks for your continued cooperation.

Read More

Answer

Thank you for providing that information.

I am sorry that it seems that this kit is not performing as expected. It would be helpful for me if you could provide me with the lot numbers or order numbers of the kits which you have found to work without any problems. If you do not have this information any more could you please estimate when these other kits were ordered?

I am contacting the lab in order to try to understand what may be contributing to this problem. I will get back to you with more information as soon as I can.

Read More

Answer

Thank you for contacting us and reporting the problems you have been encountering with the EGF Human ELISA Kit (ab100504). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

If you don't mind, could I ask a few questions in order to understand the situation a bit better.

1. How had the standard been handled prior to use? How long before carrying out this experiment was it reconstituted and how was it stored between reconstitution and testing?

2. Were the reagents allowed to cone to room temperature prior to use?

3. After which step did the precipitate begin to form? How quickly did it form?

4. Was there less precipitate with less standard present (i.e. was there no precipitate in the 0 standard well)?

5. Did you observed any precipitate in the HRP-streptavadin concentrate? Was this thoroughly mixed prior to use?

6. At what temperature was the HRP-streptavadin incubated with the plate? Was this performed in the light?

7. Was the TMB substrate reagent incubated in the dark?

I am sorry for all the questions but hopefully this information will help me to understand why this precipitate is forming. If you could also provide me with the order number and the lot number of the kit that would be very useful.

I look forward to receiving your reply.

Read More

Answer

Thank you for your enquiry. I am sorry to confirm that  Bovine samples have not been tested and guaranteed with the ab100504 EGF Human ELISA Kit,  so there is no cross reactivity data.  Epitope mapping was also not performed so we do not know exactly where the antibodies are binding.   However, we do know that the antibodies in this kit recognize the mature form of EGF. The human and bovine EGF reportedly share between 59% and 66% homology.  This percentage is not that high and it is likely the antibody would not detect in bovine samples, or would detect very weakly. The capture antibody is a monoclonal and the detection antibody polyclonal. I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Question
Answer

Thanks for your call tyesterday and for your patience while I have been in touch with the lab. I have heard back about your enquiry. Bovine samples have not been tested with the kit so we do not have any cross-reactivity data to conclusively determine this. Epitope mapping was also not performed so we do not know exactly where the antibodies are binding. However, we do know that the antibodies in this kit recognize the mature form of EGF and it looks like human and bovine EGF reportedly share between 59% and 66% homology. We would not predict the antibodies to cross-react with such low homology between the proteins.   I hope this information is useful, but if you have any further questions please let me know and I'll be happy to help.  

Read More

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

Sign up