Contents of the Kit
Protein A Cartridge tube............. 1 tube Washing Buffer .......... 13 ml x 1 bottle
Elution Buffer .............. 1.8 ml x 1 tube Catching Buffer ..........1 ml x 1 tube
1.5 mL Microtube .............. 5 tubes x 2
Storage Condition : 0-5ºC
Shipping Condition : ambient temperature
Required Equipment and Materials
Microcentrifuge, 100-200 μl adjustable pipettes, voltex mixer, 0.5 ml microtubes
IgG Purification Kits are used for isolation and purification of immunoglobulin G of goat, mouse, rabbit, and other animals. The kit contains immobilized protein A or G and buffer solutions for maximum recovery of IgG. The total time to isolate and purify IgG from serum or other solutions containing IgG is about 30 min (Fig. 1). Since the support of protein A or G is silica-base gel, the volume of the retained solution on the gel after the centrifugation is very small. Therefore, all proteins or other materials that do not bind to protein A can be removed by washing twice. SDS-PAGE of purified IgG from various animals are shown in Fig. 2. Additionally, denaturing of IgG during the elution process is minimal because IgGs bound to the gel are released with a very quick process. The gel in this kit can be used repeatedly 20 times or more with an equal performance. Used gel is also stable for one year in Washing buffer at 0-5ºC .
Fig. 1 IgG isolation process
Fig. 2 SDS-PAGE of isolated immunoglobulin by using IgG Purification
Purification Kit-A and Kit-G.
WS : Whole Serum
A : Prepared by IgG Purification Kit-A
G : Prepared by IgG Purification Kit-G
Condition: 6% acrylamide gel/Tris-glucine buffer
If the IgG solution contains gelatin,before applying the solution to the kit, enzyme digestion may be required.
Table 1 IgG Recovery from 50 μl Serum
What is the recovery of IgG with this kit?
It depends on the type of IgG and type of animal. In the case of high affinity type IgG for protein A or protein G, about 70-80% IgG is recovered from 100-200 μg IgG or IgG solution containing other proteins or macromolecules.
How is the purity of purified IgG using this kit?
The purity of the IgG from various serum is indicated in the figure above. Highly purified IgG is available with only a one time purification process.
How much IgG can be recovered from serum?
About 150-350 μg IgG can be recovered from 50 μl serum. I-7. Protein Labeling: IgG purification
How many times can a protein A gel be used?
At least 20 times.
Can I use the used protein A gel or protein G gel for the purification of different IgG solutions?
Use a new gel for a different sample to avoid contamination.
Is a used protein A gel or protein G gel stable?
A used protein A gel or protein G gel in Washing buffer is stable at 0-5ºC for one year.