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PNGase F
PNGase F can be use to cleave N-glycans attached to proteins and antibodies. This PNGase F is tag free cloned from Elizabethkingia miricola (formerly known as Flavobacterium meningosepticum).
  • Name: PNGase F
  • Synonyms: PNGase F
  • Catalogue Number:PT-EZ-PNGaseF
  • Application:Research
  • Price: 15000unit $50.00 75000unit $200.00
  • Number:
  • Concentration:50000unit/ml
  • Store:Long term storage at -20 C or below

Quality control assay: Denatured RNaseB and human monoclonal antibody were used as substrates to test PNGase F activity. (Figure above)

Stability: PNGase F retains >60% activity after left at room temperature for over 72 hours. Long term storage at - 20 C or below

Purity: >95% by SDS-PAGE gel

Unit definition: One unit is defined as the amount of enzyme required to removed >95% of the glycans from 10 ug of denatured RNase B in 1 hour at 37 C.

Concentration: 50,000 units/mL

Reference:

1. Maley, F. et al. (1989). Anal. Biochem. 180, 195-204.


Protocol for denaturing conditions

1. Denature 20 ug of glycoprotein in 0.5% SDS, 50mM DTT and 50mM Tris buffer pH8 by heating at 95 C for 5 minutes.

2. Then add NP-40 or Triton X-100 to sample (1% final concentration).

3. Add 10 uL PNGase per 20 ug of glycoprotein

4. Incubate at 37 C for 1 hour.

 

Protocol for non-denaturing conditions

1. Add 50 uL PNGase per 20 ug of glycoprotein in 50mM Tris buffer pH8

2. Incubate at 37 C for 18 hours.

 

Note:

1. Recommended substrate concentration for reaction is 0.1-1 mg/mL

2. NP-40 or Triton X-100 prevents SDS inhibition of PNGase F activity.

3. PNGase F does not cleave N-glycans containing core a1-3 fucose.

 

Cat No.: PT-EZ-PNGaseF
Product Name: PNGase F
Type: Protein
Property:
Origin:
Source: E.coli
Application: Research
Formulation: 20 mM Tris pH8, 50% glycerol.
Purity: >95% by SDS-PAGE gel
Storage: Long term storage at -20 C or below
Limitation:
Note: