HAV 004-01 is specific for an allotypic marker on human C3 (Leu instead of Pro in codon 314 of exon 9), which occurs regulsaly in the ß-chain of C3F and occasionally in the ß-chain of C3S (1). F and S mean fast and slow, respectively, on agarose gel electrophoresis. HAV 004-01 reacts with the 75-kDa ß-chain band on SDS-PAGE immunoblotting of reduced C3, and reacts with 20-kDa and 17-kDa ß-chain fragments produced by cyanogen bromide cleavage (2).
Immunogen
C3 isolated from human plasma
Application
ELISA, WB
Species reactivity (positive)
Human
Species reactivity (negative)
Not determined
Subclass
IgG2a/k
Gene ID
718
Clone number
8G4
Epitope
Epitope specificity differs from that of HAV 003-05
Content
Available in 200 µL and 1 mL size.1 mg/mL +/- 15%. See Certificate of Analysis for details.
Purification
Protein-A purified
Form
Liquid
Solvent
0.01 M phosphate buffer, pH 7.4, containing 0.5 M NaCl and 15 mM sodium azide
Storage
4-8ºC without exposure to light. No precautions necessary during handling.
Target
C3 is a key component of the complement system since classical and alternative activation pathways merge at the C3 activation step when C3 is split into C3a and C3b. The molecular mass of C3 is 185 kDa and it consists of α- and ß chains (110 kDa and 75 kDa respectively) held together by disulfide bonds (1,2).
Application 1
ELISA HAV 004-01 reacts strongly with C3. A strong reaction is seen in ELISA with C3 coated directly onto the microtiter well, and also when used as a biotinylated detection antibody in sandwich ELISA in wells coated with a polyclonal antibody against C3.
Application 2
WB
In Western blotting after SDS-PAGE, HAV 004-01 detects the above-mentioned allotypic marker on the ß-chain of C3, closely related but not completely specific to the fast-moving form of C3, C3F.
HAV 004-01 can be cleaved by papain to produce Fab fragments. (1, 2)