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choice of antibody


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#1 Zoya

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Posted 18 July 2011 - 02:06 PM

Hi all,

I am doing a tyrosine hydroxylase (TH) immunostaining experiment on Dopamine neurons in VTA. I just decided to use a fluorescence microscope, so I would need to use the fluorescent detection. I want to work with Mouse anti-human TH (primary antibody) and Horse anti-mouse IgG (secondary antibody), but my supervisor ask me to find the best antibodies to have the best results. I would appreciate if someone help me to find the right primary and secondary antibody.

Thank you so much.

#2 bob1

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Posted 18 July 2011 - 03:53 PM

The best fluorescent antibodies out there are the Alexafluor range from Molecular probes (now Invitrogen/Life technologies).

#3 Zoya

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Posted 18 July 2011 - 10:18 PM

The best fluorescent antibodies out there are the Alexafluor range from Molecular probes (now Invitrogen/Life technologies).


But my problem is the choice of primary and secondary antibodies. Do you have any suggestion for them to get the best result?

Thank you.

#4 bob1

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Posted 19 July 2011 - 03:54 PM

I know nothing about TH antibodies, so I can't help you there, other than to suggest that you look at the literature and see what other people have used.

Once you have that sorted, look on the Invitrogen website for the Alexafluor antibody that corresponds (e.g. goat anti mouse alexafluor488 conjugate) to your primary...

The numbers after the antibody correspond to the excitation spectrum of the antibody, so 488 is excited by blue light with a maximum of 488 nm and emits green light.

#5 centropagus

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Posted 14 December 2012 - 12:50 PM

At 26 kDa, GST is considerably larger than many other fusion protein affinity tags. For reasons that have not been fully characterized in the literature, the structure of the GST fusion tag often degrades upon denaturation and reduction for protein gel electrophoresis (e.g., SDS-PAGE). As a result, electrophoresed samples often appear as a ladder of lower MW bands below the full-sized fusion protein.




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