Overview

Description

  • Nature
    Native
  • Source
    Native

Specifications

Our Abpromise guarantee covers the use of ab9936 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Biological activity
    SDS PAGE profile of a typical recombinant Troponin-I preparation (3.8ug) showed a major band at 24kDa and a minor band attributable to Troponin-I dimer. Please note: Troponin-I is very susceptible to proteolytic breakdown within cardiac tissue and it is difficult to isolate undegraded native human cardiac troponin-I. This product is greater than 70% intact and hence proteolytic fragments can be observed.
  • Form
    Liquid
  • Additional notes
    Native Human Cardiac Troponin-ISDS PAGE profile of a typical recombinant Troponin-I preparation (3.8ug) showed a major band at 24kDa and a minor band attributable to Troponin-I dimer. Please note: Troponin-I is very susceptible to proteolytic breakdown within cardiac tissue and it is difficult to isolate undegraded native human cardiac troponin-I. This product is greater than 70% intact and hence proteolytic fragments can be observed.
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.

    pH: 8.00
    Constituents: 48% Urea, 0.117% Beta mercaptoethanol, 0.79% Tris HCl, 0.0292% EDTA

General Info

  • Alternative names
    • cardiac muscle
    • Cardiac troponin I
    • cardiomyopathy, dilated 2A (autosomal recessive)
    • Cardiomyopathy, familial hypertrophic, 7, included
    • CMD1FF
    • CMD2A
    • CMH7
    • cTnI
    • Familial hypertrophic cardiomyopathy 7
    • MGC116817
    • RCM1
    • Tn1
    • Tni
    • TNN I3
    • TNNC 1
    • TNNC1
    • TNNI3
    • TNNI3_HUMAN
    • Troponin I
    • Troponin I cardiac
    • Troponin I cardiac muscle
    • Troponin I cardiac muscle isoform
    • Troponin I type 3 cardiac
    • troponin I, cardiac 3
    • TroponinI
    • Ttroponin I type 3 (cardiac)
    see all
  • Function
    Troponin I is the inhibitory subunit of troponin, the thin filament regulatory complex which confers calcium-sensitivity to striated muscle actomyosin ATPase activity.
  • Involvement in disease
    Defects in TNNI3 are the cause of cardiomyopathy familial hypertrophic type 7 (CMH7) [MIM:613690]. Familial hypertrophic cardiomyopathy is a hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death.
    Defects in TNNI3 are the cause of cardiomyopathy familial restrictive type 1 (RCM1) [MIM:115210]. RCM1 is an heart muscle disorder characterized by impaired filling of the ventricles with reduced diastolic volume, in the presence of normal or near normal wall thickness and systolic function.
    Defects in TNNI3 are the cause of cardiomyopathy dilated type 2A (CMD2A) [MIM:611880]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
    Defects in TNNI3 are the cause of cardiomyopathy dilated type 1FF (CMD1FF) [MIM:613286]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
  • Sequence similarities
    Belongs to the troponin I family.
  • Information by UniProt

Images

  • Natural Cardiac Troponin I protein (ab9936) at 2.7 µg (Coomassie blue staining of SDS gel)
    Performed under reducing conditions.

    Observed band size: 24 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 48 kDa (possible dimer)

References

ab9936 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase. This product is listed as "This peptide can be used with studies usinghttps://www.abcam.com/index.html?datasheet=7829,https://www.abcam.com/index.html?datasheet=8290andhttps://www.abcam.com/index.html?datasheet=8291.", all of which list ELISA as a tested application. If you are having difficulty using this in ELISA I would be happy to help. If you could send me your protocol and any data that you may have, I could look over that and if possible offer suggestions which may help. If we are unable to solve this problem I would be more than happy to send ab50222 as a replacement for ab9936.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Application
Sandwich ELISA
it works well. However it gets very sticky in high concentrations

Abcam user community

Verified customer

Submitted Dec 08 2014

Answer


Troponin I is a very hydrophobic protein, that is why it either precipitates or sticks to glass/plastic ware in the absence of carrier proteins.

In order to avoid the precipitation, Troponin I can be prediluted with rabbit skeletal muscle troponin-C solutions. Troponin C complexes with Troponin I, renders it soluble and, to some extent, protects it from protease activity

Troponin-I is very susceptible to proteolysis, the urea buffer confers stability to TnI because it denatures contaminating proteases. Therefore, we cannot guarantee the TnI stability when diluted with other buffers.

The addition of 8M urea could indeed cause some trouble when using antibodies. I am pleased to present you an alternative product that does not contain urea. Human Cardiac Troponin I full length protein (ab50803), this is a recombinant full length protein and its constituents are 0.5M Sodium chloride, 60mM Beta mercaptoethanol, 20mM Tris HCl, pH 7.5. More information here:

https://www.abcam.com/index.html?datasheet=50803 (or use the following: https://www.abcam.com/index.html?datasheet=50803).

Human Cardiac Troponin I full length recombinant protein (ab50803) is guaranteed by our Abpromise (www.abcam.com/abpromise) for SDS-PAGE and WB.

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Answer

I have been in contact with the lab who have confirmed that ab9936 is pure troponin-I and it does not contain Troponin-C. I would try and use this as the standard.
ab9919 is pure skeletal muscle troponin-I (no troponin-C) and therefore would be good to use as a control.
Development of stable TnI controls is a very difficult process. The lab usually adds TnI to a 25-fold excess of TnC, incubates until recovery stabilizes (up to several days), and lyophilizes. Stability is also very dependent on the epitopes recognized by the antibodies used in a particular assay. For that reason, it is not possible to provide a procedure that will work for everyone.

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Answer

Troponin-I can only be purified in urea containing buffers. In the absence of urea, at concentrations above ug/ml it precipitates and/or sticks to glass or plastic war. Even in the presence of 9 M urea it completely retains its functional ability to bind calcium dependently to troponin-C. When complexed to troponin-C, troponin-I is soluble. The complexed form is also much more resistant to protease degradation. For these reasons, troponin-I standards usually consist of TnI complexed to TnC.
The urea buffer also confers stability to TnI because it denatures contaminating proteases.

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Answer

Thank you for contacting us.
We have following anti Cardiac Troponin IgGs.
https://www.abcam.com/cardiac-troponin-i-antibody-ab47003.html
https://www.abcam.com/cardiac-troponin-i-antibody-284-19c7-ab19615.html
https://www.abcam.com/cardiac-troponin-i-antibody-ep1106y-ab52862.html
https://www.abcam.com/cardiac-troponin-i-antibody-4c2-ab10231.html
https://www.abcam.com/cardiac-troponin-i-antibody-ab155047.html
The anti Troponin I are
https://www.abcam.com/skeletal-muscle-troponin-i-antibody-42-25-14b-ab97427.html
https://www.abcam.com/troponin-i-fast-slow-skeletal-muscle-antibody-12f10-ab110132.html
Please carefully review the datasheets and buy the product as required.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you very much for your call. As discussed I am happy to send you this confirmation:

Abcam takes product quality seriously and I would like to reassure you that we investigate all customer complaints.In addition to our in housequality control, we monitor customer feedback closely on a weekly basis and we have received no complaints about this product. Thus, we are not currently concerned about the general qualityof the Cardiac Troponin I protein ab9936 when shipped on this conditions.

We ship this protein in packaging with ice packs to provide extra temperature stability in transit. The ice pack may be thawed or even at room temperature when you receive it; please be assured that this is normal and that your product is safe to use in the tested and guaranteed applications (i.e. in Western blot/SDS-PAGE). Once you have received the vial, please follow the long-term storage instructions on the datasheet, i.e. storage at -80ºC and avoid freeze/thaw cycles. Also, the storage buffer including 8M urea should ensure suitable conditions for the protein to be used as stated on the datasheet.

Our Abpromise to you is that, should our product not work as stated on the datasheet, we will resolve the issue to your satisfaction, either with helpful advice to optimize your experiment or a replacement or refund of the product. Please contact our knowledgeable Scientific Support staff should you experience any issues while using our products. Contact information can be found on our website ('Contact us', or use mailto:technical@abcam.com).

I hope this information helps, but please don't hesitate to contact me for additional information or assistance.

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Answer

Thank you for your reply.

Further investigation into this protein revealed that this is very tightly bound and hydrophobic (http://www.rcsb.org/pdb/explore/explore.do?structureId=1MXL). As such the full peptide will need to be stored and used in a reduced form to be effective. Ab50822 is a single strand peptide and do not require these conditions.

While using this in ELISA is not ideal, the protein structure itself makes this a necessity. A shorter incubation at room temperature, one hour, with the sample may provide better results.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Answer

Thank you for contacting Abcam.

I would not recommend using a product in ELISA with a storage buffer which contains8 M Urea, 15 mM ß-mercaptoethano as I would be wary of the buffer denaturing your antibody. I would recommend using a peptide which has a non-reducing buffer such as ab50822;https://www.abcam.com/cardiac-Troponin-I-protein-ab50822.html.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Question
Answer

Thank you for contacting us.

The purity is determined by observation of a stained SDS-PAGE gel. The information I received from our source of the cardiac troponin protein is as follows:

Purity is estimated visually from the SDS PAGE gel. It probably exceeds 95%. Troponin-I is very susceptible to proteolysis and lower mwt bands on SDS PAGE represent TnI fragments (confirmed by western blot). The proteolysis occurs during the time the heart is in cold storage. With human hearts, it is extremely difficult to obtain human hearts that have been frozen immediately after death.

Please do not hesitate to contact us if you need any advice or more information.

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1-10 of 14 Abreviews or Q&A

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