Anti-cardiac Troponin I antibody [2D5] (ab7829)

Thank you for your helpful reply. Regarding Cardiac Troponin I protein ref. ab9936, I see that it has been only tested for WB. Do you have information using it for ELISA? In other case, do you have a specific one for ELISA?  Regards, 

ANSWER:

Thank you for your email.

The ab9936 product is a peptide which can be used to block antibodies ab7829, ab8290 and ab8291. This peptide can be used to block these antibodies irrespective of applications e.g. It is suitable to use with antibodies in Immunoprecipitation, western blot, Immunohistochemistry, ELSIA & also other blocking assays.

I hope this information will be helpful. Should you have any other question please do not hesitate to ask.

I'm interested in proving cardiac differentiation of P19 cells on the protein level. I'm wondering if you have antibodies (best monoclonal) specific for cardiac muscle cells respectively unspecific for skeletal muscle cells, e.g. cardiac troponin I/T/C, cardiac MHC, cardiac actinin, ...

There seem to be some on your website such as cardiac TnI (ab7829), however, i'm not sure if there is no cross-reactivity with skeletal muscle. Can you help? Many thanks for your efforts!!

ANSWER:

Thank you for your enquiry.

I looked at our cardiac troponin antibodies and found several by using our browser's filter function. The reactivity of these antibodies with skeletal troponin is listed on the datasheets; on some datasheets it is listed in the Specificity section and on others in the Relevance section.

For P19 cells, you want to use an antibody that has been tested on mouse, or an antibody that has been generated using an immunogen that is 100% identical to the homologous mouse sequence. I found ab19615, ab10237 (troponin I) and ab10214 (troponin T). Ab7829 is also non-reactive with skeletal troponin but has only been tested on human tissue.

There may be more troponin antibodies that will work for you, depending on your application. The filter allows you to search by application as well as by species reactivity and by species the antibody was raised in. I hope what I have suggested is a helpful start. If you have any other questions, please do not hesitate to contact us.

Does ab7829 cross-react with skeletal muscle troponin I? What is the percentage cross-reaction?

ANSWER:

Thank you for your enquiry.

Our supplier states that there will be no cross reactivity with skeletal muscle troponin I. They do not have any number for percentage cross-reactivity though.

I'm using your product ab9935 (cardiac Troponin I) and its antibody ab7829 they work well in western blotting I have a question? The MW of about 24kDa is given not only by cardiac Troponin I, I suppose, but probably by another protein you used to precipitate the E.coli product (for example GST): wich is this other protein? and which are the single MW of the two protein?

ANSWER:

Thank you for your enquiry.

There is no tag on ab9935. It was purified using Troponin I affinity chromatography. The MW of about 24kDa is given only by cardiac Troponin I.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

I've performed a Western Blotting just to test Cardiac troponin I protein (#ab9935) and and Cardiac troponin I antibody (#ab7829) I used your protocol with Ab incubation at 4°C overnight: I obtained more than 1 band!!!!!! I can see 6 bands, but in particulare3: the biggest one (for size and density) is at about33 KDa the second (for size and density) is between 18 and 26 KDa the third (for size and density) is smaller than 18 KDa.

Is it normal? and what is the Cardiac troponin I protein? How is it possible with a purified protein and a specific Ab from the same customer? If you can justify it, ho can I justify referee?

I've bought everithing because the in vitro translation I did was dirty and Ab anti tags too aspecific and referee wants something clearer but now I've this problem...

ANSWER:

I have just received the following information from the source of ab9935:

"The recombinant troponin-I is indeed 95% pure or greater. It was prepared from E. coli by TnC chromatography in urea which pretty much guarantees purity. Recombinant proteins are sometimes expressed partially translated or proteolytically degraded and the resultant fragments will purify with the intact polypeptide. The western results tell me that the Ab you are using also recognize an epitope on the low level TnI fragments. Reactivity on westerns rarely correlates with staining intensity on SDS PAGE. Sample preparation in reducing buffers is ideal for TnI."

We have added an image of the SDS-PAGE separation of the protein ab9935 on the datasheet, you can observe the band band size at 24kDa with an additional band at around 48kD which represents a possible dimer.

I hope this information helps, please do not hesitate to contact us again if you need further assistance,