Since these proteins are always undergoing recycling, I would suggest using GFP transgenic animals to study the dynamics through live imaging. However if that is not possible, I believe there is a Rab4 antibody commercially available that you can use. I am not sure if there is an antibody for Rab11 yet.
It depends on which recycling are you studying. There are various recycling routes, and this paper explains them very clearly http://www.nature.com/nrm/journal/v5/n2/full/nrm1315.html .
If you are looking for something possibly related to the early endosomes and not to the recycling from Golgi, I suggest to use anti-transferrin receptor antibody. it is bright in immunofluorescence and it marks ALL the recycling pathway that involves both Rab11 and Rab4. Instead, I strongly discourage the use of Rab antibodies, even if various antibodies are available in the market. They are really shitty in my hands, very low fluorescence and difficult interpretation of the results. In addition to transferrin receptor, also anti-EEA1 antibodies work well (early endosomes) .
If you need some other suggestion, don't hesitate to contact me.
good luck!
I agree with Chiara, anti-transferrin receptor, or fluorescently labeled transferrin are the best tools available when following recycling routes via immunofluorescence.
wo endosome populations involved in recycling of membranes and receptors to the plasma membrane have been described, the early and the recycling endosome. However, this distinction is mainly based on the flow of cargo molecules and the spatial distribution of these membranes within the cell. To get insights into the membrane organization of the recycling pathway, we have studied Rab4, Rab5, and Rab11, three regulatory components of the transport machinery. Following transferrin as cargo molecule and GFP-tagged Rab proteins we could show that cargo moves through distinct domains on endosomes. These domains are occupied by different Rab proteins, revealing compartmentalization within the same continuous membrane. Endosomes are comprised of multiple combinations of Rab4, Rab5, and Rab11 domains that are dynamic but do not significantly intermix over time. Three major populations were observed: one that contains only Rab5, a second with Rab4 and Rab5, and a third containing Rab4 and Rab11. These membrane domains display differential pharmacological sensitivity, reflecting their biochemical and functional diversity. We propose that endosomes are organized as a mosaic of different Rab domains created through the recruitment of specific effector proteins, which cooperatively act to generate a restricted environment on the membrane.
Keywords: endocytosis, transferrin recycling, Rab proteins, EEA1
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