Loose Resin
Protein G resin is formulated in 20% ethanol. Do not freeze the resin (freezing the suspension will damage the agarose beads), or store it at room temperature. All resins are susceptible to oxidative agents and high temperatures should be avoided. The resin is resistant to short exposure of 8M urea, pH 11 and pH 1.0. Protein G is resistant to treatment with 0.1M NaOH.
Specifications
| Protein G |
Recombinant Protein G expressed in E. coli deficient in albumin binding domain |
| Supporting matrix: |
4% crosslinked agarose resin |
| Ligand Density: |
2 mg Protein G/ml resin |
| Bead size range: |
45-165 μm |
| Recommended working pH: |
pH 2.5- 9.0 |
| Typical binding capacity: |
Upto 20 mg Human IgG /ml resin |
| Linear Flow rate |
Upto 300 cm/h (5cm diameter column, pressure 1 bar) |
| Maximum Flow rate |
20-40 ml/min |
| Optimum Flow rate |
1-10 ml/min |
| Maximum pressure |
0.1MPa (1 bar) |
| Chemical stability: |
High |
| Solubility in water: |
Insoluble |
| Toxin Levels |
Free of Staphyloccocus enterotoxins and hemolysins |
Detemine Antibody Concentration
For pure solution the Beer-Lambert law, A = ε.c.l, can be used to determine the protein concentration of IgG (mg/ml).
Extinction Coefficient (ml.mg-1.cm-1)
| IgG |
0.72 |
| IgM |
0.84 |
| IgA |
0.94 |
Sandwich ELISA assay can also be used to accurately measure antibody concentrations within a range of 1 mg/ml to 20 mg/ml sample. The antibodies can also be monitored for purity by SDS-PAGE under reducing or non-reducing conditions (see page xxx). Note that IgG appears in a reducing SDS-PAGE as 25 kDa and 50-55 kDa bands and IgM appears as 25 kDa and 70-80 kDa bands. Recovery of immunoglobulins can be quantified by Bradford assay (see pages 14 & 15), scanning densitometry of reducing or non-reducing SDS-polyacrylamide gels or ELISA.
IDT
