Hi Suhana, I use multiplex for cytokine detection. I can really recommend it. You will have a absolute quantification of your protein concentration. I have not experience with the levels of these cytokines in the bronchoalveolar fluid. But I guess during allergy you will have enough for its detection.

Hi all, thanks for your response and information. I think I'll consider using this kit.

Hi, I started using the Vplex from MSD to detect cytokines in unconcentrated human BAL last which was impossible a few years ago. I also used luminex to measure cytokines in concentrated Bal. the levels were low. If you got MSD platform in your university. It is best to use Vplex from MSD because they are more sensitive than other multiplex system I used today.

Hi, you have to make sure you concentrate the BAL fluid so you can detect cytokines. freeze drying has proved to be a very good opotion in our hands. The miliplex kits from BioRad or Millipore are very good.

Hi, it should work but it's important to concentrate you samples. I use the milliplex kits from Millipore and they works good. Good luck.

hi, thanks for your suggestions. As for concentrating the BAL fluid, what method should I use other than freeze drying? I've read that some people used Amicon centrifugal filter from Milipore to concentrate protein sample, can I use that filter for BAL fluid? 

Hi Suhana, the Luminex platform works fine and I would recommend it rather than e.g. the CBA kit from BD. I am not familiar with bronchoalveolar lavage fluid, but please keep in mind that concentrating the BAL fluid might interfere with your assay performance (LLoD etc.). You should evaluate this before using your clinical samples by spiking and concentrating control BAL with chosen cytokines and then analyse the recovery rate. By the way, classical ELISA are still more sensitive than multiplex bead assays. If you have only a few specific cytokines in mind you should consider using them instead. In case of overall screening of specimen multiplex assays are your best option, of course.

Regards, Werner

Hi,

i used a lot of the multiplex kits from Bio-rad and some of the luminex kits from R&D but for me the Bio-rad kits worked better because of their concentration range for my cytokines. The R&D kit was ok but the range was lower in each direction (lowest and highest concentration range) and even for a standard curve the Bio-rad kits had 2 more dilution steps (in my case better because i measured some cytokines in very low concentrations which would be lost with the R&D kits).

I haven´t tested Millipore yet but know from a colleague that they worked for her too.

Oh and Bio-rad is more expensive if thats important for your choice.

Hi,

The kit works good. But if you are working with low concentrations (I have no experience with bronchoalveolar fluid), try to include more dilutions for your standard. Only pay attention at the MFI values, your background should not be lower than your last dilution. 

 Hi Salme Suhana,

I am aslo interested in performing luminex assays using Mouse BAL fluid, and I was wondering if the R&D kit worked for you.

if so, what dilution did you use for your BAL samples?

I appreciated any suggestions from you regarding the assay

thanks

Hello Salme

I am planning to use Luminex in BAL samples, can you share witch cytokines you measured and the dilutions you used?? how did it work?

If you don't have MSD platform in your institution, contact the company (www.mesoscale.com) and send your samples to them to measure for you. They have very sensitive assays in sub pg levels.

Could anyone recommend me a specific kit/tool to concentrate BAL samples and associated protocol? I'm struggling to find information online, thanks