When we use Polyclonal antibody to detect a virus using western blotting， two kinds of virus particles were found ，while the virus we used have never been reported such case， I do not think this is Defective interfering particles (DIPs) ,I want to konw why？
whether other virus have situation like this？
Thank you.
I agree with Guenther. The most important question I would have is: What virus is it, which cell culture (or specimen) does it come from and how was the antiserum from which animal established?
More Information is needed. Which virus, how purified, gele electrophoresis conditions etc...
I agree with Guenther. The most important question I would have is: What virus is it, which cell culture (or specimen) does it come from and how was the antiserum from which animal established?
Dear Liu Yong-xiang
I Agree With Guenther Keil and Sven M. Bergmann. several factors affecting virus multiplication, extraction, purification...?
Regards
Prof. Houda Kawas
Dear Liu Yong-xiang,
I agree with the other colleagues, we need to know, which virus is in use. The viruses are extremely different in their properties. Just two examples:
1) When working with the RNA-containing lymphocytic choriomeningitis virus (LCMV), we performed exact measurements using highly purified virus particles, which were divided into infectious units (IU) and interfering particles (IP) containing a shortened genome. IU were calculated as plaque-forming units (PFU) and mouse IU, where the titers of the latter were in general tenfold higher. Counting total particles via electron microscopy and comparing them with titrations revealed ratios for PFU of 1:20, (i.e. 1 IU in 20 particles) and for IFU of 1:4000 (i.e. 1 IP in 4000 particles).
2) Later I was concerned with the DNA-containing duck hepatits B virus (DHBV), which was a mixture of infectious particles with so-called subviral particles (SVP) lacking a genome at all. Here, purification and separation techniques of those entities disclosed a strong surplus of SVP over infectious particles. This could easily followed when viral genome equivalents (vge) of infectious particles were compared with the large surface proteins (L) of both particles. The calculations uncovered that purified infectious particles consisted of 3.9x10E9 vge and 4.1x10E10 L, whereas purified SVP consisted of
influenza may be in spheroid or filamentous forms, depending on culture conditions
Hello,
In this case these are not two different viral particles.may be single protein existing in doublet or little non specific since poly ab. you can use monoclonal and try the same.
All the best
Dear Liu Yong-xiang,
Western blotting detecte proteins of viruses not viral particles. I see only two proteins at this blotting picture. May be it's two proteins of one virus. May be it's contamination. I agree with the other colleagues we need to know more about your material and methods.
Best regards
These bands are HA protein, ~60 kDa. We see the same picture after WB of H7N9 viruses with mouse sera raised against whole virus. May be the protein is in two forms - glycosylated and non-glycosylated?
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