~42kDa. Due to glycosylation, PTX3 may migrate between 40-45kDa.
Source:
Produced in CHO cells. Human full-length PTX3.
UniProt ID:
P26022
Concentration:
0.1mg/ml.
Formulation:
Liquid. In PBS containing 2% BSA and 0.1% sodium azide.
Applications:
ELISA, WB
Application Notes:
ELISA and Western blot standard together with MAb to PTX3 (Prod. No. ALX-804-464 (capture antibody)) and PAb to PTX3-Biotin (Prod. No. ALX-210-365B (detection antibody)).
Shipping:
Shipped on Blue Ice
Long Term Storage:
-20°C
Handling:
Avoid freeze/thaw cycles.
Protocol:
Sandwich ELISA for PTX3
96 well-ELISA plates (Nunc MaxiSorp 446612) were coated with 100µl of MAb to PTX3 (human) (MNB4) (Prod. No. ALX-804-464), 1μg/ml in 100µl coating buffer (15mM carbonate buffer pH 9.6) and incubated overnight at 4°C.
After incubation, the plates were washed three times with 300µl/well of washing buffer (PBS + 0.05% Tween 20), and then 300µl of 5% dry milk in washing buffer were added to block non-specific binding.
The plates were incubated for 2 hours at room temperature and then washed three times with washing buffer.
50µl (in duplicate) recombinant human PTX3 standard (Prod. No. ALX-201-124) (75pg/ml to 2.4ng/ml ), cell supernatants or biological fluids diluted in PBS without Ca and Mg + 2% BSA were added and then incubated for 2 hours at 37°C.
The plates were washed five times with washing buffer, and then 100µl of PAb to PTX3 (human) (Biotin) (Prod. No. ALX-210-365B) 50ng/ml in washing buffer were added in each well.
The plates were incubated for 1 hour at 37°C and then washed five times with 300µl of washing buffer.
100 µl/well streptavidin-horseradish peroxidase diluted 1:4’000 were added and the plates incubated for 1 hour at room temperature.
After incubation the plates were washed 5 times with washing buffer and 100µl of TMB substrate solution was added to each well. After incubation for 10 min. at room-temperature, 50μl of stop-solution (1M H2SO4) was added. Subsequently, absorbance was read within 30min at 450nm.
Note: Do not use heparin plasma or serum samples with this assay. EDTA samples are recommended. The detection value in serum is generally less than in plasma samples.
Regulatory Status:
RUO - Research Use Only
Product Literature References
Antitumor and anti-inflammatory effects of trabectedin on human myxoid liposarcoma cells: G. Germano, et al.; Cancer Res. 70, 2235 (2010), Abstract;
The long pentraxin Ptx3 is synthesized in IgA glomerulonephritis and activates mesangial cells: B. Bussolati, et al.; J. Immunol. 170, 1466 (2003), Abstract;
Long pentraxin PTX3 upregulates tissue factor expression in human endothelial cells: a novel link between vascular inflammation and clotting activation: E. Napoleone, et al.; Art. Thromb. Vasc. Biol. 22, 782 (2002), Abstract;
Non-redundant role of the long pentraxin PTX3 in anti-fungal innate immune response: C. Garlanda, et al.; Nature 420, 182 (2002), Abstract;
Circulating levels of the long pentraxin PTX3 correlate with severity of infection in critically ill patients: B. Muller, et al.; Crit. Care Med. 29, 1404 (2001), Abstract;
Dynamic induction of the long pentraxin PTX3 in the CNS after limbic seizures: evidence for a protective role in seizure-induced neurodegeneration: T. Ravizza, et al.; Neuroscience 105, 43 (2001), Abstract;
PTX3 in small-vessel vasculitides: an independent indicator of disease activity produced at sites of inflammation: F. Fazzini, et al.; Arthritis Rheum. 44, 2841 (2001), Abstract;
PTX3, A prototypical long pentraxin, is an early indicator of acute myocardial infarction in humans: G. Peri, et al.; Circulation 102, 636 (2000), Abstract; Full Text
The long pentraxin PTX3 binds to apoptotic cells and regulates their clearance by antigen-presenting dendritic cells: P. Rovere, et al.; Blood 96, 4300 (2000), Abstract; Full Text
Expression of a long pentraxin, PTX3, by monocytes exposed to the mycobacterial cell wall component lipoarabinomannan: V. Vouret-Craviari, et al.; Infect. and Immunity 65, 1345 (1997), Abstract;
Multimer Formation and Ligand Recognition by the Long Pentraxin PTX3. Similarities and differences with the short pentraxins C-reactive protein and serum amyloid P component: B. Bottazzi, et al.; J. Biol. Chem. 272, 32817 (1997), Abstract; Full Text
