€1,750.00
GlyCLICK conjugation of MMAE
GlyCLICK contains all reagents and materials needed to label IgG with GlyCLICK conjugation of DBCO-VS(GlcA)-PAB-MMAE.
Antibody Conjugation, ADC Development
GlyCLICK
Cloned from S. pyogenes, Produced in E. coli (His-tagged), Engineered galactosyltransferase (Y289L)
Enzymatic components: Immobilized GlycINATOR (EndoS2) on agarose beads (20% EtOH), beta-1,4-galactosyltransferase (GalT)
IgG Fc-glycans
GlycINATOR hydrolyzes Beta-1,4-linked GlcNAc residues on all IgG Fc-glycoforms for site-specific azide activation using GalT and complete labeling using SPAAC
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
Deglycosylation (GlycINATOR): 120 min incubation at 37°C, Azide Activation (GalT): O/N incubation at 30°C, Labeling (SPAAC): O/N incubation at 25°C, *The protocol can vary depending on kit size/amount of IgG
TBS (pH 7.4)
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).
GalT requires buffer additive for activity., Buffer additive is included in the GlyCLICK kit.
Denaturated IgG
Components for antibody conjugation of up to 0.25-2 mg IgG
MMAE with Cleavable Linker (Val-Ser(GlcA)-PAB)