Exoquick

How do I decide which ExoQuick product to start with? ExoQuick or ExoQuick-TC??

ExoQuick should be used with biofluids that are concentrated in exosomes like serum, plasma, or ascites fluid. ExoQuick-TC should be used with biofluids that are dilute in exosomes like tissue culture (cell culture) media, urine, and CSF.

Can my biological fluid have been frozen before using ExoQuick?<

Frozen biofluids should be thawed on ice and then proceed with the regular ExoQuick protocol.

What are the typical yields of exosome, protein and RNAs from various biological fluids?

See the values in the Table below. These are average yields.

Biological Fluid	Starting Volume	Number of Particles	Amount of Protein	Amount of RNA
Serum/ Plasma	250 ul	6 x10^11	10 mg	250 ng
Urine (typical)	5 ml	5 x10^9	50 ug	25 ng
Tissue Culture Media	10 ml	This depends on the type of cells that were grown in culture and how many exosomes they secrete.

When I used ExoQuick, a large insoluble pellet was obtained and now it is difficult to work with. What should I do?

Typically this happens if starting with plasma or serum. With plasma, there are a lot of fibrins and fibrinogens that also get precipitated with ExoQuick. We recommend adding the Thromboplastin D protocol before adding the ExoQuick, per the steps in the user manual. For serum, this typically occurs if you have incubated the ExoQuick with the serum for too long, or if the centrifugation step was too long. See the troubleshooting table below.

Biological Fluid	Preparation steps	ExoQuick product to use	Volume of Fluid: ExoQuick	Incubation Time	Centrifugation Speed/ Length of Spin
Serum	None	ExoQuick	4:1 (v:v)	30 minutes	13,000 rpm/ 2 min
Plasma	Thromboplastin D treatment. See supporting protocol in User Manual (link)	ExoQuick	4:1 (v:v)	30 minutes	13,000 rpm/ 2 min

What should I resuspend the precipitated exosomes with?

This depends on what you want to do with the exosomes. Follow the guidelines here:

Further Application	Buffer to use
Western Blotting	RIPA buffer
ExoELISA	Exosome Binding Buffer
RNA isolation	Lysis Buffer for RNA extraction kit
Exosome transfer studies	Water, PBS, DMEM, or other media

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