Anti-ss DNA antibody [TNT-3] (ab78856)

How much DNA can be detected using these antibodies? Do you have information on levels of detection? i.e. how much DNA can be detected per ug of antibody?

ANSWER:

Thank you for your inquiry and I appreciate your patience with this reply.
For ab27156,the lab did nothave any data on how much DNA can be detected with the ds DNA antibody. However different specificities regarding the binding characteristics of the antibody are described below:
Specificity: Primarily double stranded DNA (dsDNA). Measurements by immuno-CE yielded KD’s of 0.71 μM and 0.09 μM, for the interaction of ab27156 with ss- and dsDNA, respectively (1). Strong reactivity with both ss- and dsDNA has been observed on dot blots as well as very weak reactivity with RNA.
Reactivity:In combination with immunofluorescence the antibody is well suited for detection of dsDNA in fx. Crithidia luciliae, a monoflagellate protozoa, containing a giant mitochondrion (2). The antibody can be used for easy quantification of in vitro cell proliferation by ELISA and has successfully been used for monitoring cytotoxity and apoptosis as well as ELISA-based co-culture angiogenesis and proliferation assays (3). Very good reactivity both with dsDNA and ssDNA on NC-dotblots has been observed. The minimal size for DNA bindingab27156 is >16 bases and there is an inverse proportionality between binding and ionic strength (1).
1) N.H.H. Heegaard, D.T. Olsen and K.P. Larsen (1996) Journal of Chromatography A, 744: 285-294. (attached)
2) J.B Peter and Y. Schoenfeld (1996) Autoantibodies, Elsevier. .230-236.
3) T. Friis, B. Kjaer Sorensen, A.M. Engel, J. Rygaard and G. Houen (2003) APMIS. 111(6):658-68.
For ab78856, the lab did not have any further data on the detection limits of the antibody andno information on the amount of DNA detected per ug of the antibody.
I hope this information helps. Please contact us with any other questions.

How much DNA can be detected using these antibodies? Do you have information on levels of detection? i.e. how much DNA can be detected per ug of antibody?

ANSWER:

Thank you for your inquiry.

I'm trying to find more information for you from the lab, but in the meantime here are the ELISA references we talked about on the phone for ab78856

Hornick, J.L., et al.. Cancer Biotherapy & Radiopharmaceuticals, 13:255-268 (1998).

Khawli, L.A. et al.. Cancer Biotherapy & Radiopharmaceuticals , 17:359-370 (2002).

I'll be in touch once I have more information.

I recently bought your antibody (Anti-ss DNA antibody [TNT-3] (ab78856)) for detection of ssDNA...
I have a question belowon this antibody...

Which part of DNA (ssDNA and dsDNA) can be recognized by this Abcam anti-ssDNA antibodies?...

If this antibody binds at the end of DNA, very small number of antibodies only bind to DNA in case of using whole genomic DNA... if this is true, I think shearing DNA or fragmentation of DNA would be better because after shearing, many DNA fragments are recognized by the antibodies...

However, if this is not true, please answer me to my inquiry above...
Thank you very much and have a good day...

ANSWER:

Thank you for contacting Abcam about ab78856.

The antibody only recognizes single stranded DNA but not double stranded DNA. There is no specific information on what part of a ssDNA will be binding to so I cannot answer the question about shearing DNA. However, that would be a non natural condition of your samples so that won't reflect the actual amount of ssDNA in the samples. If you want to see NA you can do a stain with a DNA specific dye.
The following article explains how the antibody was developed, it may be of interest for you: http://www.ncbi.nlm.nih.gov/pubmed/10850361

Please let me know if there is anything else I can help you with.

Are there any more details available for the protocol used to test this antibody in ELISA?

ANSWER:

Thank you for your call yesterday and for your patience while I have been in touch with the lab regarding your enquiry.

This antibody was tested in ELISA by an academic collaborator in the following article:

Hornick, J.L., et al.. Cancer Biotherapy & Radiopharmaceuticals, 13:255-268 (1998).

Khawli, L.A. et al.. Cancer Biotherapy & Radiopharmaceuticals , 17:359-370 (2002).

Unfortunately I don't have access to these articles so I don't have further information regarding the ELISA protocl used. I would recommend starting around 0.1 ug/mL as we discussed, and optimizing from that point.

I am sorry that I don't have more information, but if you have any questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.