Overview

  • Product name
    Human Sonic Hedgehog ELISA Kit
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 20 pg/ml
  • Range
    13.72 pg/ml - 10000 pg/ml
  • Recovery

    94 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 91.72 82% - 101%
    Serum 95.34 84% - 103%
    Plasma 76.43 68% - 89%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

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

    This assay employs an antibody specific for Human Sonic Hedgehog coated on a 96-well plate. Standards and samples are pipetted into the wells and Sonic Hedgehog present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Sonic Hedgehog 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 Sonic Hedgehog bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Optimization 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
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    Assay Diluent C 1 x 30ml
    Biotinylated anti-Human Sonic Hedgehog (lyophilized) 2 units
    Recombinant Human Sonic Hedgehog Standard (lyophilized) 2 vials
    Sonic Hedgehog Microplate (12 x 8 wells) 1 unit
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas
  • Relevance
    Sonic Hedgehog, which is expressed only during embryogenesis, is instrumental in patterning the early embryo. It has been implicated as the key inductive signal in patterning of the ventral neural tube, the anterior-posterior limb axis, and the ventral somites. Of three human proteins showing sequence and functional similarity to the Sonic Hedgehog protein of Drosophila, this protein is the most similar. The protein is made as a precursor that is autocatalytically cleaved; the N-terminal portion is soluble and contains the signalling activity while the C-terminal portion is involved in precursor processing. More importantly, the C-terminal product covalently attaches a cholesterol moiety to the N-terminal product, restricting the N-terminal product to the cell surface and preventing it from freely diffusing throughout the developing embryo. Defects in this protein or in its signalling pathway are a cause of holoprosencephaly (HPE), a disorder in which the developing forebrain fails to correctly separate into right and left hemispheres. HPE is manifested by facial deformities. In addition, it is thought that mutations in this gene or in its signalling pathway may be responsible for VACTERL syndrome, which is characterized by vertebral defects, anal atresia, tracheoesophageal fistula with esophageal atresia, radial and renal dysplasia, cardiac anomalies, and limb abnormalities.
  • Alternative names
    • HHG 1
    • HHG1
    • HLP3
    • HPE3
    • MCOPCB5
    • SHH
    • SMMCI
    • Sonic hedgehog homolog
    • Sonic hedgehog protein
    • Sonic hedgehog protein C product
    • Sonic hedgehog protein N product
    • TPT
    • TPTPS
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab100639 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

  • Representative standard curve using ab100639

  • Representative standard curve using ab100639

Protocols

References

This product has been referenced in:
  • Estep M  et al. Hepatic sonic hedgehog protein expression measured by computer assisted morphometry significantly correlates with features of non-alcoholic steatohepatitis. BMC Gastroenterol 19:27 (2019). Read more (PubMed: 30744560) »
  • Ihnatovych I  et al. Timing of Wnt Inhibition Modulates Directed Differentiation of Medial Ganglionic Eminence Progenitors from Human Pluripotent Stem Cells. Stem Cells Int 2018:3983090 (2018). Read more (PubMed: 30050576) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Answer

Unfortunately, I can not find a reference for using our Human ShhN ELISA kit with serum or plasma.

However, I would like to reassure you that we have successfully validated this kit with serum and plasma samples ourselves and therefore this product is covered by our abpromise guarantee.

Read More
Abreviews
We tested this product on pig brain tissue (thawed from snapfrozen in liquid nitrogen), since we were informed that there is a 95% homology between human and pig SHH protein sequences. Since no information was available for which diluent to use, we found diluent B most appropiate and used this. We followed the protocol step by step.
We had no problem getting acceptable results from the ELISA and we will now proceed to order yet another kit for the remaining brain samples to be analyzed.
Although the kit worked fine, the manual is somewhat messy at places (e.g. the peculiar dilutions used in the standard curve could use some explanation), but nevertheless, the kit seems to work fine in pig brain tissue.

Abcam user community

Verified customer

Submitted Nov 21 2013

Question
Answer

Thank you very much for your calls and for your patience while we have been in touch with the lab regarding your enquiry.

Both the capture and detection antibodies are monoclonal, and as expected the epitopes have not been mapped so we don't know exactly where the antibodies target. We do know that the antibodies bind theN-termius (aa 23-198)but most of the N-terminus is similar to the Indian hedgehog protein so cross-reaction is likely.

I am sorry that we don't have more information at this time, but if you have further questions or need anything else, please let me know and I'll be happy to help.

Read More

Question
Answer

Thank you for contacting Abcam regarding ab100639.


Assay Diluent B would be more appropriate if testing amniotic fluid (or any other body fluid other than serum or plasma).


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More

Answer

Thank you for contacting us. The constituents of the diluents are proprietary, but I am fairly certain you will want to use diluent C for amniotic fluid. I have asked the laboratory to confirm and will forward their reply.

Read More

Answer

Thank you for your reply.

The antibody pair is considered intellectual property so they are not provided separately from the kits. Could you describe the exact assay that you would like to use the antibodies in?Ican ask the lab again and see if there is anything else that we can do in this situation, but I am notconfident that it will be possible.

I am sorry for this inconvenience,but I look forward to hearing from you. Please let me know if you have any further questions.

Read More

Question
Answer

Thank you again for contacting us and for your patience.

Unfortunately the lab is not able to provide these antibodies as components separate from the kit ab100639. I am very sorry for the inconvenience.

We do have a pair of sonic hedgehog antibodies that have been used in a sandwich ELISA with a recombinant human SHH fragment as the analyte, ab19897 and ab87382-

https://www.abcam.com/sonic-hedgehog-antibody-ab19897.html

https://www.abcam.com/sonic-hedgehog-antibody-10h6-ab87382.html

Please let me know if you have any questions or if there is anything else we might be able to do for you, and I'll be happy to help.

Read More

Question
Answer

Thank you very much fory our call yesterday and for your interest in these antibodies.

I am still working on getting some more information about these antibodies, and I thank you for your patience. Our office will be closed tomorrow, but I will be back in touch with you on Thursday.

Please let me know if you have further questions or if there is anything else that we can do for you. Have a nice day.

Read More

Question
Answer

Thank you for contacting Abcam regarding ab100639.


We are not able to provide the exact amino acid sequence of this protein standard. This is the N-terminus amino acid sequence for the Sonic Hedgehog protein. I can tell you that the predicted molar mass is 20 kDa.


I hope this information is somewhat helpful. Please do not hesitate to contact me if you have any additional questions.

Read More

Answer

Thank you for your inquiry.

As Assay Diluent B does contain proteins and other solutes it would not be abnormal to see a little cloudiness or precipitation, especially after being stored at a low temperature and/or for a long period of time.

The lab tested Diluent B that showed precipitation and I can confirm that there are no adverse affects. We recommend to spin the Diluent Bdown and take just thesupernatant for the experiments. Alternatively, theDiluent B can be filtered.

Please note that while these aggregates may look visual unappealing, based on evaluated customer data and feedback, they do not seem to affect the assay. Nevertheless, the lab is aware that these aggregates are unsettling to customers and is taking further steps to reduce their appearances.

I hope this information is helpful. Please do nothesitate contact me again.

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1-10 of 20 Abreviews or Q&A

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