Protein G HP SpinTrap
Protein G HP SpinTrap is a prepacked, single-use spin column containing Protein G Sepharose High Performance.
FAQ
The products can be used for two application areas:
- enrichment of proteins/immunoprecipitation/pulldowns/immunoaffinity, several different terms are used for this type of preparation—enhancing the signal of a protein of interest from clarified cell lysates and biological fluids
- small-scale purification of antibodies—screening of clones or the purification of small quantities of antibodies
For details on the usage of the specific products, see the table below.
| Product | Protein enrichment | Antibody Purification |
| Ab SpinTrap | X | X |
| Ab Buffer Kit | X | |
| NHS HP SpinTrap | X | |
| NHS HP Buffer Kit | X | |
| Protein A HP SpinTrap | X | X |
| Protein A HP MultiTrap | X | X |
| Protein A/G HP SpinTrap Buffer Kit | X | |
| Protein G HP SpinTrap | X | X |
| Protein G HP MultiTrap | X | X |
| Streptavidin HP SpinTrap | X | |
| Streptavidin HP MultiTrap | X | |
| Streptavidin HP Buffer Kit | X |
Reproducible enrichment of native proteins minimizes variation in sample preparation for quantitative and comparative expression studies. Standardized protocols optimized for downstream analysis such as electrophoresis and LC-MS. High enrichment capability ensures improves downstream detection. Includes protocols that support performance optimization, modification and troubleshooting
Immunospecific affinity sample preparation procedures are based on the interaction between an antibody or similar molecule and its’ ligand protein. This is a platform for seven protein enrichment products with the flexibility of coupling an already established affinity molecule of a wide variety of types and also from a broad range of species. The affinity molecule on the column will be used as a bait to capture and enrich the proteins of interest in the sample with high specificity. The majority of the other proteins in the sample will be eliminated in flow through and wash fractions of the columns.
What is the pH stability of Protein G Sepharose High Performance?
pH 3-9 (long term), 2-9 (short term).
Note that a pH below pH 3 is sometimes required to elute strongly bound antibody species. However, protein ligands may hydrolyze at very low pH.
What molecule is immobilized ?
Protein A and Protein G HP SpinTrap and MultiTrap:
Immobilization of antibodies from a wide variety of sources. The relative binding strengths of different subclasses of an antibody from various species differ. See Table below for more information.
Relative binding strengths of antibodies from various species to protein G as measured in a competitive ELISA test. Antibody binding strength to protein A is also shown for comparison. Cytiva also offers prepacked Protein A HP SpinTrap and Protein A HP MultiTrap; see Ordering information for details
| Species | Subclass | Protein G binding | Protein A binding |
| Human | IgA | — | variable |
 |
IgD | — | — |
|
IgE | — | — |
|
IgG1 | ++++ | ++++ |
|
IgG2 | ++++ | ++++ |
|
IgG3 | ++++ | — |
|
IgG4 | ++++ | ++++ |
|
IgM* | — | variable |
| Avian egg yolk | IgY† | — | — |
| Cow | |
++++ | ++ |
| Dog | |
+ | ++ |
| Goat | |
++ | — |
| Guinea pig | IgG1 | ++ | ++++ |
|
IgG2 | ++ | ++++ |
| Hamster | |
++ | + |
| Horse | |
++++ | ++ |
| Koala | |
+ | — |
| Llama | |
+ | — |
| Monkey (rhesus) | |
++++ | ++++ |
| Mouse | IgG1 | ++++ | + |
|
IgG2a | ++++ | ++++ |
|
IgG2b | +++ | +++ |
|
IgG3 | +++ | ++ |
|
IgM* | — | variable |
| Pig | |
+++ | +++ |
| Rabbit | |
+++ | ++++ |
| Rat | IgG1 | + | — |
|
IgG2a | ++++ | — |
|
IgG2b | ++ | — |
|
IgG3 | ++ | + |
| Sheep | |
++ | +/— |
* Purified using HiTrap™ IgM Purification HP columns, code no. 17-5110-01
† Purified using HiTrap IgY Purification HP columns, code no. 17-5111-01
++++ = strong binding
++ = medium binding
— = weak or no binding
What is Protein G HP SpinTrap and Protein G HP MultiTrap?
Protein G HP SpinTrap is 16 prepacked spin columns packed with Protein G Sepharose High Performance medium. Protein G HP MultiTrap is a 4 pack with prepacked 96-well filter plates packed with Protein G Sepharose High Performance. Protein G Sepharose High Performance medium enables immobilization of IgG antibodies with high affinity. Antibodies are immobilized only or with the option to covalently couple the antibody via cross-linking to Protein G ligand and the protein/antigen of interest is enriched from the sample. The protein of interest can be enriched up to several hundredfold depending on the specificity of the antibody.
How do I store Protein G HP SpinTrap and Protein G HP MultiTrap?
Store at 4°C to 30°C in 20 % ethanol
How are the SpinTrap columns used?
SpinTrap columns are prepacked, single use spin columns that are used one by one or in parallel together with a bench top centrifuge for enrichment of proteins of interest.
Are different protocols available for Protein G HP SpinTrap/Protein A HP SpinTrap and Protein G HP MultiTrap/Protein A HP MultiTrap?
Yes, It’s a choice between classic protocol for speed or a cross-link protocol ensuring only elution of the antigen and not the antibody.
What is the matrix for Protein G Sepharose High Performance?
The matrix is a highly cross-linked spherical 6% agarose, with the ligand recombinant protein G (lacking albumin-binding region) coupled with NHS-activation. The mean particle size is 34 μm.
What is the difference between Protein G HP SpinTrap and Ab SpinTrap?
The product Protein G HP SpinTrap and Ab SpinTrap are exactly the same columns. The difference is the application and the protocols supplied with the products. Protein A HP SpinTrap and Protein G HP SpinTrap protocols are developed for enrichment of selected target proteins. First the antibody is immobilized to the Protein G and secondly the protein/antigen of interest is enriched from the sample.
Ab SpinTrap columns are designed for rapid small-scale purification of the antibodies themselves. The antibody binds to the Protein G ligand, and then the column is washed to remove unbound material before the antibody is eluted.