Dear Wasyl, 

Plz do Real time PCR for all. Especially in the case of MMP, you can run Native PAGE and Do densitometric studies or Image analyzer 

i have estimate those tnf-alpha and mmp,ete in college level.

we dont have such facility like pcr and elisa.

for that i have to estimate by using biochemical methods so i need a solution madam @ Katarzyna Wasyl

dear murugan sir

i have to estimate inflammatory mediators from asthamatic/copd lung, so i need  any biochemical methods for estimation of those inflammatory mediators other than ELISA AND PCR

Hi, if you are looking in extra-cellular fluids you can use FACS beads coated with anti-mouse IgG Abs and specific mouse Abs against your target molecules.

hi wasyl madam,

ya,but now am at college so we  don't have that so....

Maybe you can try these: http://www.rndsystems.com/product_results.aspx?c=220&s=0&o=AND&a=0&cj=-1

These membranes have antibodies against many different inflammatory mediators immobilized in them. You don't need any specialized equipment to run the assay, excepting a dark room and x-ray film in the final step.

I have already tried them and they are an amazing tool.

The pros

• easy to use
• fast assay
• detection of multiple proteins simultaneously

The cons:

• expensive
• limited number of samples (4 membranes in each kit)

Hope it helps

Multiplex assays me be useful for your purposes.

Can you do cellular assays, like cell killing assays or ones that are responsive to TNF alpha.  For proteases, you can use FRET substrates with media from cells  ( MMPs and ADAMs).  For caspases, many companies sell kits. 

thanks one and all