PNGase F (Peptide N-glycosidase F)
N-Glycan을 가수분해하는 PNGase F 효소가 결합되어 있는 Spin column 타입으로, 짧은 반응시간이 장점이며, MS Analysis에서 Enzyme peak 없이 분석 가능합니다.
Deglycosylation Workflow
항체, Fusion Protein 등 N-당화 당단백질의 N-Glycan을 가수분해함
Robust and handles both native and denaturing conditions
2 개의 포맷으로, Native 와 Denaturing condition 모두 적용 가능
빠른 반응 시간
Denaturing conditions: 15분+
Native conditions: 1시간~16시간(Overnight)
PNGase F 효소가 Resin에 결합되어 있는 Spin Column 형태
N-Glycan을 가수분해한 후 Centrifugation을 통해 순도 높은 Deglycosylated protein 획득
MS analysis에서 Enzyme peak 감소 효과
Unit Definition
0.05 ml of settled agarose resin hydrolyzes ≥ 95% of N-glycans from 0.2 mg etanercept in TBS, pH 8.6 in 1 h at 37°C as monitored by SDS-PAGE.
Buffers
The recommended buffer for best performance of the column is TBS buffer at pH 8.6. Other buffers with neutral pH containing 0.15 M NaCl can also be used but optimization may then be required.
Content and Storage
Immobilized PNGase F Microspin columns contain sufficient material to remove N-glycans from 0.2 mg glycoprotein. The resin is supplied in 20% EtOH with no preservatives added, and are provided in packs of 5 or 10 columns. The columns are shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the product!
Immobilized PNGase F Denaturing contains microspin columns of Immobilized PNGase F (see above) and vials of RapiGest™* SF Surfactant. One 1 mg RapiGest™ SF vial is included per column of Immobilized PNGase F. RapiGest™ SF should be stored at room temperature. Once reconstituted in high purity water or a buffer (pH 7–10), the solution is stable for one week when stored at +2–8 °C.
| Part number | Item name | Units | Description |
| G1-PF6-010 | Immobilized PNGase F | 5 x 0.2 mg | Microspin type |
| G1-PF6-020 | 10 x 0.2 mg | ||
| G2-PDK-010 | Immobilized PNGase F Denaturing | 5 x 0.2 mg | |
| G2-PDK-020 | 10 x 0.2 mg |