GlyCLICK Azide Activation

825.005,145.00

GlyCLICK Azide Activation enables site-specific conjugation of IgG using a label of choice

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Description

GlyCLICK Azide Activation is a kit that contains all reagents needed to azide-activate IgG for site-specific conjugation using any alkyne-carrying label of choice. Spin columns with GlycINATOR (EndoS2) enzyme immobilized on agarose beads deglycosylates IgG from several species and subclasses by hydrolyzing any Fc glycoform after the inner most GlcNAc. The GalT enzyme supplied in solution enables incorporation of the UDP-GalNAz regent to the exposed GlcNAc sugar for subsequent click chemistry incorporating any alkyne-carrying label of choice. GlyCLICK Azide Activation is available in kit formats to facilitate activation of 250 ug, 2 mg or 10 mg of IgG.

Detailed Information

Application

Antibody Conjugation, ADC Development, Custom Conjugation

Product Group

GlyCLICK

Source/Origin

Cloned from S. pyogenes, Produced in E. coli (His-tagged), Engineered b-1,4-galactosyltransferase (Y289L)

Product Composition

Enzymatic components: Immobilized GlycINATOR (EndoS2) on agarose beads (20% EtOH), beta-1,4-galactosyltransferase (GalT)

Digestion Site

IgG Fc-glycans

Specificity

GlycINATOR hydrolyzes Beta-1,4-linked GlcNAc residues on all IgG Fc-glycoforms for site-specific azide activation using GalT

Substrates

Human IgG1, Human IgG2, Human IgG3, Human IgG4, Cow IgG, Fc-fusion proteins, Goat IgG, Horse IgG, Monkey IgG, Mouse IgG, Rabbit IgG, Rat IgG, Sheep IgG

Reaction Conditions

Deglycosylation (GlycINATOR): 30 min incubation at 37°C, Azide Activation (GalT): O/N incubation at 30°C, *The protocol can vary depending on kit size/amount of IgG

Digestion Buffers

TBS (pH 7.4)

Label

No label

Buffer Compatibility

GlyCLICK steps are performed in TBS buffer, provided in the kit., For purification of azide activated Ab, TBS or PBS can be used., Sodium azide interferes with the click reaction and must be avoided., Avoid phophate buffers, not compatible with buffer additive (precipitation).

Additives/Co-factors

GalT requires buffer additive for activity., Buffer additive is included in the GlyCLICK kit.

Inhibitors

Denaturated IgG

Unit Definition

Components for antibody azide activation of up to 0.25-10 mg IgG

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