N-Glycan Label A0212

Description: N-Glycan Label is used in ultra-fast labeling of glycans of PNGase F glycosidase-digested N-linked glycoproteins

 (including antibodies). Labeled glycan has highly sensitive flfluorescence characteristics and ionization properties by 

mass spectrometry, allowing for quantitative analysis using LC-FLR and LC-MS. The unique chemical properties of this 

label provide higher fluorescence quantification and mass spectrometry response.

Physical Appearance: Lyophilized powder

Molecular Weight: 427.1850 Da

Molecular Formula: C21H25N5O5

Mass Shift: + C17H20N4O2 = 312.3663 Da 

Labeled N-Glycan (Da) = Released N-Glycan + Glycan Label Reagent - Reaction by-product ( C4H5NO3, MW: 115.0268) - HH

Resuspension: Dissolve 4.5 mg N-Glycan label in 55 μL freshly prepared DMF solution to reach 82 μg/μL.

Storage Conditions: -20 °C refrigerator as lyophilized powder.

Shelf Life: 12 months at -20 °C.

Application: 4.5 mg N-Glycan labeling reagent can label fifive digested glycan samples.

Reference Procedure: 

1. Add 10 μL desalted 1.5 μg/μL mAb samples into 0.6 mL low protein binding collection tube.

2. Add 10 μL 3% BT Surfactant with 5 mM TCEP and 50 mM HEPES (pH 7.9) to the tube and mix well. 

3. Heat at 95 °C for 3 min, then cool to the room temperature.

4. Add 10 μL 1.5 U/μL rPNGase F and mix well.

5. Incubated at 50 °C for 15 - 30 min, then cool to room temperature.

6. Add 10 μL 82 μg/μL Glycan Label, incubate at room temperature for 5 min.

7. Add 15 μL anhydrous DMF and 45 μL acetonitrile. Mix well.

Labeled N-glycan samples can be used for LC-FLR and LC-MS analysis.

Product Composition: The ratio of N-Glycan Label: NHS compound is 3:2 analyzed by LC-QE HF.

Labeling Efficiency: 99.5% when 0.37 μg/μL mAb and 20 μg/μL N-Glycan tags are used to label 15 μg N-glycans released

 by glycoproteins.

Thermal stability: N-Glycan labeling reagent is stable in anhydrous DMF at room temperature for 8 h; Labeled glycoamines

 are stable at room temperature for 24 h, and can withstand 320 °C of mass spectrometry ionization temperature.

Chemical Instability: Unstable when concentrations of affinity reagents, SDS, Tris, DTT, amine, and mercaptans greater 

than 0.1 mM.