PNGase F is a glycosidase that catalyzes the cleavage of internal glycosidic bonds in oligosaccharides. PNGase F removes almost all N-linked oligosaccharides from glycoproteins. PNGase F releases N-glycans from glycoproteins during glycoanalysis workflows.

Physicochemical Properties

Molecular Formula	C21H38O3
Molecular Weight	338.52462720871
Exact Mass	2123.698
CAS #	83534-39-8
PubChem CID	167676231
Appearance	Colorless to light yellow liquid
Hydrogen Bond Donor Count	8
Hydrogen Bond Acceptor Count	39
Rotatable Bond Count	8
Heavy Atom Count	155
Complexity	3670
Defined Atom Stereocenter Count	0
InChi Key	UVVLNLNAGIIHGJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H12FN3O3.C13H11N3O4.2C13H13N3O3.C12H10N4O4.3C12H12N4O3/c1-6-4-15-11-7(10(6)14)5-17(13(11)20)8-2-3-9(18)16-12(8)19;1-6-4-7-10(14-5-6)13(20)16(12(7)19)8-2-3-9(17)15-11(8)18;1-7-4-8-9(14-5-7)6-16(13(8)19)10-2-3-11(17)15-12(10)18;1-7-4-8-6-16(13(19)11(8)14-5-7)9-2-3-10(17)15-12(9)18;1-5-4-13-8-9(14-5)12(20)16(11(8)19)6-2-3-7(17)15-10(6)18;1-6-13-4-7-8(14-6)5-16(12(7)19)9-2-3-10(17)15-11(9)18;1-6-4-13-10-7(14-6)5-16(12(10)19)8-2-3-9(17)15-11(8)18;1-6-4-13-7-5-16(12(19)10(7)14-6)8-2-3-9(17)15-11(8)18/h4,8H,2-3,5H2,1H3,(H,16,18,19);4-5,8H,2-3H2,1H3,(H,15,17,18);4-5,10H,2-3,6H2,1H3,(H,15,17,18);4-5,9H,2-3,6H2,1H3,(H,15,17,18);4,6H,2-3H2,1H3,(H,15,17,18);4,9H,2-3,5H2,1H3,(H,15,17,18);2*4,8H,2-3,5H2,1H3,(H,15,17,18)
Chemical Name	6-(2,6-dioxopiperidin-3-yl)-3-methylpyrrolo[3,4-b]pyrazine-5,7-dione;6-(2,6-dioxopiperidin-3-yl)-3-methylpyrrolo[3,4-b]pyridine-5,7-dione;3-(4-fluoro-3-methyl-7-oxo-5H-pyrrolo[3,4-b]pyridin-6-yl)piperidine-2,6-dione;3-(3-methyl-5-oxo-7H-pyrrolo[3,4-b]pyrazin-6-yl)piperidine-2,6-dione;3-(3-methyl-7-oxo-5H-pyrrolo[3,4-b]pyrazin-6-yl)piperidine-2,6-dione;3-(3-methyl-5-oxo-7H-pyrrolo[3,4-b]pyridin-6-yl)piperidine-2,6-dione;3-(3-methyl-7-oxo-5H-pyrrolo[3,4-b]pyridin-6-yl)piperidine-2,6-dione;3-(2-methyl-5-oxo-7H-pyrrolo[3,4-d]pyrimidin-6-yl)piperidine-2,6-dione
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

Suitable for: 1. De-N-glycosylation of glycoproteins such as immunoglobulin fusion proteins, antibodies, or other in vitro; 2. determination of whether proteins have undergone N-glycosylation modification. PNGase F stock solution contains 10 mM Tris (pH7.4), 50 mM NaCl, and 5 mM EDTA. Molecular weight: 36 kDa. Ideal digestion temperature: 37°C. pH range: 6.0-10.0. Temperature range: 4-50°C. Ionic strength range: 0-1 M NaCl. Because glycerol is absent from the stock solution, it is simpler to adjust chromatographic conditions for HPLC techniques. Reaction Buffer (10X): Sodium Phosphate, 500 mM, pH 7. N-glycosides containing core α1-3 Fucose, which is commonly present in plant and insect glycoproteins, cannot be hydrolyzed by PNGase F; PNGase A is therefore necessary.

References

[1]. Evaluation of different PNGase F enzymes in immunoglobulin G and total plasma N-glycans analysis. Glycobiology. 2021 Jan 9;31(1):2-7. [2]. Highly Efficient Release of Glycopeptides from Hydrazide Beads by Hydroxylamine Assisted PNGase F Deglycosylation for N-Glycoproteome Analysis. Anal Chem. 2015 Oct 20;87(20):10199-204.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.9540 mL	14.7702 mL	29.5404 mL
	5 mM	0.5908 mL	2.9540 mL	5.9081 mL
	10 mM	0.2954 mL	1.4770 mL	2.9540 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.