FAQ
Which are the features and benefits of using Albumin & IgG Depletion SpinTrap?
1. Improved detection of lower abundant proteins in human plasma/serum
2. Simple and fast procedure
3. High depletion capacity, removes > 95% HSA and > 90% IgG, with high reproducibility
4. High sample loading capacity
5. Short protocol
6. Flexible usage through multiple formats; use SpinTrap with a tabletop centrifuge and HiTrap with an automated LC system or manually with a syringe
How is Albumin & IgG Depletion SpinTrap stored?
Albumin & IgG Depletion SpinTrap is stored filled with 20 % ethanol at + 2°C to + 8°C.
What are HiTrap Albumin & IgG Depletion and Albumin & IgG Depletion SpinTrap?
These products are prepacked columns used for the depletion of albumin and IgG from human plasma and serum samples.
What is the material used in SpinTrap?
Polypropylene barrel and polyethylene frits
How do you operate SpinTrap?
SpinTrap is operated using a standard tabletop centrifuge
Which samples can the depletion products be used for?
The products have been optimized for human plasma and serum, but can in theory also be used for other body fluids such as cerebrospinal fluid where it may be beneficial to deplete albumin and IgG. Tissue samples rich in blood where albumin and IgG can interfere with the analysis have not been tested.
Can the Albumin & IgG Depletion SpinTrap column be re-used?
Reusing the Albumin & IgG Depletion SpinTrap has not been evaluated, thus we can only recommend single-use.
Who are the target customers?
All customers that work with protein research analyzing human plasma or serum samples who are interested in analyzing proteins of less abundance, and do not have an affinity-based identification and detection method. Customers can be found within drug discovery departments in a pharmaceutical company or within academic or medical research, with areas of interest such as biomarker research, proteomics, and protein biochemistry.
When is depletion useful and what are the main applications?
High abundant protein depletion adds value to any analysis when a high resolution of the ingoing proteins is required, and when the aim is to analyze a number of proteins and no specific detection is available for the individual proteins. Examples of these types of analyses are 2-D electrophoresis/DIGE.
What is the mechanism of the depletion process?
The sample is added to the column, and albumin and IgG is bound to the matrix. The flow-through contains the depleted sample.
How does high abundant protein depletion add value?
Serum and plasma contains a very high concentration of proteins and is also a quite common sample type as it relatively easy to obtain. However, it is also a quite challenging sample, in that the dynamic range of ingoing proteins is very large. By far, the most abundant proteins are albumin and IgG, which together make up approximately 70% of the total protein content
of the sample. The two main reasons that depleting the sample of these proteins will add value to the analysis are:
1) Less abundant proteins can then be detected since the signals from albumin and IgG very often obscure the signal of a large portion of the protein content in the sample.
2) By removing these high abundant proteins, a relatively higher amount of less abundant proteins can be subjected to the analysis, thus increasing the sensitivity of the analysis.
When is the SpinTrap format useful?
Using the SpinTrap format is very useful when a smaller volume of sample is being processed, and when a number of samples will be processed in parallel.
What is the capacity and performance of the Albumin & IgG Depletion SpinTrap column?
A load of 50 ìl plasma containing normal levels of albumin and IgG (~40 mg HSA/ml and ~15 mg IgG/ml) results in > 95% albumin depletion and > 90% IgG depletion.
What is the Albumin & IgG Depletion SpinTrap?
It is a spin column filled with approximately 370 ìl agarose-based matrix with affinity ligands for human albumin and IgG. It is designed for the depletion of albumin and IgG from human plasma or serum.
What will happen in the depletion step to the proteins that are bound to albumin and IgG in the sample?
Proteins bound to albumin and IgG when the sample is applied will commonly stay bound to these molecules, and thus will also be depleted from the sample. The bound fraction can, however, be eluted from the column and analyzed separately.
What media and matrix is prepacked into the HiTrap Albumin & IgG Depletion and Albumin & IgG Depletion SpinTrap?
Both columns are prepacked with a mixture of anti-HSA Sepharose High Performance and Protein G Sepharose High Performance. The mixed medium is based on a highly cross-linked agarose with an average particle size of 34 ìm. The ligand for anti-HSA Sepharose High Performance is a recombinant protein with affinity for human albumin, and the ligand for Protein G Sepharose High Performance contains protein G fragment with an affinity for IgG.