The Thermo Scientific Pierce Albumin Depletion Kit uses an agarose resin of the affinity ligand Cibacron* Blue dye for high-capacity, selective extraction of human serum albumin from 10 to 50μL serum samples.
Pierce Albumin Depletion Resin is supplied as a 50% slurry. Dispense 200μL of slurry into the supplied microcentrifuge spin columns to obtain 100μL of settled beads for the standard protocol. Each aliquot of resin can be used to process 10 to 50μL of serum sample in a single reaction. Processed samples are ready for immediate downstream analysis by electrophoresis or MS applications. The ease and versatility offered by the slurry format make this kit ideal for single- or multi-sample processing with the microcentrifuge spin columns supplied in the kit.
- Complete kit – includes optimized buffer and microcentrifuge spin columns to remove albumin quickly and conveniently from 10 to 50μL samples
- Easy-to-dispense slurry – improves performance, eases processing of multiple samples, and can be adapted to larger or smaller columns or 96-well filter plates
- Workflow compatible – removing excess albumin facilitates MS or electrophoresis gel analysis of low-abundance serum proteins
- Resin slurry, binding-wash buffer, and microcentrifuge spin columns
Human serum albumin (HSA) often accounts for greater than 60% of the total protein present in serum samples and can have a concentration of approximately 40mg/mL. The high concentration of albumin hinders research, obscuring the detection of many proteins of biological interest. Traditionally, researchers have produced albumin-free samples using chromatography methods involving multiple purification steps. In addition to involving lengthy and tedious procedures, these purification steps also tend to give low protein yields. The Pierce Albumin Depletion Kit was developed to take advantage of the Cibacron* Blue Dye albumin binding properties.
||Effective albumin removal improves 2D gel analysis of serum. A. Human serum was diluted five-fold in TBS (i.e., 10μL of serum added to 40μL of TBS), and 5μL of the diluted serum was separated by 2D-SDS PAGE. B. Human serum was diluted two-fold in Binding/Wash Buffer (i.e., 50μL of serum added to 50μL of buffer) processed using the Pierce Albumin Depletion Kit according to the instructions. Albumin-depleted samples and washes were combined (i.e., 100μL sample with 150μL wash buffer) and 5μL was separated by 2D-SDS PAGE. For 2D-SDS PAGE analysis, samples were diluted with 2D SDS-PAGE loading buffer, focused using pH 4-7 isoelectric focusing (IEF) strips and separated using 8-16% Tris-glycine gels. Gels were stained using GelCode Blue Stain (Part No. 24590). The black arrow points to the gel region where albumin is located on the duplicate gels before (A) and after (B) processing.
The kit is optimized to bind human, porcine and sheep albumin from serum samples. With a modification to the protocol, albumin from bovine, calf, goat and rat can be removed with this method. The kit is not effective for removal of mouse albumin.
Review of affinity purification
Pierce Albumin/IgG Removal Kits
Abundant Protein Depletion Spin Columns