MagicLink™ HRP Antibody Conjugation Kit
HRP 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 amount of homo-crosslinking. The thiol-maleimide linkageis
exchangeable when other thiol groups are present. In the 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 an issue.
A typical flow chart of HRP -antibody conjugation by MagicLink technology
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 HRP to yield antibody-HRP conjugate with a stable ring linkage.
· 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 HRP 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
· HRP activity: > 250 U/mg
MagicLink HRP antibody Labeling Kits are used to conjugate HRP with primary antibody, secondary
antibodies, and other biomolecules for the bioassay below.
· Western blotting (WB)
· Immunohistochemistry - Paraffin
· ELISA Capture
· ELISA Detection
· Flow cytometry
· Immunohistochemistry - Frozen
· MAGIC NHS (MW ~900)
· LINK activated HRP
· Reaction buffer
· Protein concentrator
Instruction sheet (PDF)