Specificity
Binds to human high
molecular weight single-chain urokinase type plasminogen activator
(HMW-scuPA, 54 kDa), to high molecular weight two-chain urokinase
type plasminogen activator (HMW-tcuPA, 52 kDa) and to low molecular
weight single-chain urokinase type plasminogen activator (LMW-scuPA,
33kDa) with high affinity (Kd = 0.1 nM) and does inhibit the ability
of urokinase to activate plasminogen. Urokinase reaction with
synthetic substrate S2444 is not inhibited. Highly specific for
urokinase, no cross-reactions were found with other serine proteases
(tissue plasminogen activator, plasmin, trypsin, chymotrypsin,
elastase, and thrombin). Human plasma or plasma proteins do not
interfere with its binding to urokinase.
Preparation
Precipitation by PEG-6000
and DEAE-cellulose chromatography (gradient)
Format
Lyophilized from 0.01M sodium phosphate, 0.01M sodium borate, 0.15M sodium chloride, 1% dextran, 1%mannitol, pH 8
Applications and
recommended dilutions
Immunohistochemistry
(Frozen Sections), Enzyme Immunoassay (EIA), Functional Studies
(Func), Western blotting (WB). Can be used for detection of urokinase
(RIA, ELISA), affinity purification and for studies on the structural
and kinetic parameters of the enzyme. Can be used in immunoblotting
(Western, dot-blot) and immunohistochemistry on frozen sections. The
antibody is very stable and can, after appropriate binding, be used
20-40 times in immunopurification of urokinase in 1 M guanidine
chloride. For immunohistochemistry, dilute approximately 1:30 - 1:50
and for ELISA, dilute starting from 1:5,000. For Western blotting
dilute 1:100. Other applications not tested. Optimal dilutions of
this antibody are dependent on conditions and should be determined by
the user.
Reconstitution
Use 1 ml sterile pure
water
Storage
Shipping at ambient
temperature. Storage of lyophilized antibodies for 2 years at –20°C
or lower
