MagicLink™ Alkaline Phosphatase Antibody Labeling Kit

A typical flow chart of AP-antibody conjugation by MagicLink™ technology.

Alkaline phosphatase (AP) can be conjugated to antibody/protein by thiol-maleimide and hydrazone

coupling approaches. The thiol-maleimide approach is commonly performed with a bi- functional linker

(such as SMCC). However, SMCC-modified protein is extremely unstable and often self-reactive since it

often contains both amine and thiol groups that cause significant amounts of homo-crosslinking. The thiol-

maleimide linkage is exchangeable when other thiol groups are present. Inthe hydrazone coupling approach,

the conjugation reaction is based on a reaction between hydrazine and aldehyde. This method is low

yielding although a catalyst is used. The size of the molecule and inaccessibility of the functional group

contribute to low yield. In addition, linkage stability is a concern.

BroadPharm's MagicLink™ technology solves all the problems by combining PEG and proprietary

crosslinking chemistry. MagicLink™chemistry and kit do not need TCEP, DTT, or catalysts at all!

First, antibody reacts with MAGIC NHS ester to yield the MAGIC activated antibody which instantly reacts

with LINK preactivated AP to yield antibody-AP conjugate with a stable ring linkage.

Features;

• Novelty: 3rd generation protein crosslink technology

• Yield: conjugation efficiency around 95-100%

• Rate:instant reaction; the whole process less than 1 hour

• Linkage stability: most stable covalent bonded conjugates on the market

• Consistence: good reproducibility from batch to batch

• Flexible: use your own AP and antibody at your desired ratio

• Shelf life: long, stable, freeze dried ship at ambient temperature

• Antibody selection: any proteins/antibody, antibody fragment

• Post-conjugation clean-up step not necessary

• AP activity:  4200 U/mg, high specific activity grade

Application;

Alkaline Phosphatase Labeling Kit is used mainly for the preparation of alkaline phosphatase antibodies and

other proteins in immunological techniques such as ELISA, Western blotting, and immunohistochemistry.

Consists of;

• MAGIC NHS (MW ~900)

• LINK activated AP

• Reaction buffer

• Protein concentrator

  Instruction sheet (PDF)