Depletion kits

Following are some aspects that limit the broad application of the depletion kits. The potential issue of employing this albumin depletion strategy is the limited sample size, typically less than 50 pL, which can be treated by the depletion kits. This sample size prevents the use of a large sample volume to obtain assay sensitivity enhancement. The depletion kits are relatively expensive. Perhaps the most significant drawback of the immunodepletion methods is nonspecific removal of plasma proteins of interest that may be bound to the removed proteins (Anderson and Hunter, 2006 Liu et al., 2006). 

Depletion of these and several other abundant proteins allow for in-depth analysis of low-abundance ones. There are many commercial products for protein immunodepletion. Applications of the products are reported for a Seppro column in a label-free method (135), Seppro column in a SILAP method (136), a Hu-6 Multiple Affinity Removal System (MARS) kit in an iTRAQ method (137), and a Hu-6 MARS kit in another iTRAQ -based method (138). Caution should be observed if a low-abundance protein has affinity to a high-abundance protein because it can be co-depleted. 

Figure 3. BITC-induced intracellular GSH depletion. The cells were treated with 25 pM BITC (open square) or phenyl ITC (open triangle) for different time intervals. Intracellular GSH levels were coloremetrically measured using commercial kit GSH-400 (BIOXYTECH).

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