Quantifying Residual Lectin Pathway Activity
Corresponding Organization : Mario Negri Institute for Pharmacological Research
Other organizations : University of Copenhagen, Rigshospitalet
Protocol cited in 4 other protocols
Variable analysis
- Coating material: mannan (10 μg/mL) or acetylated bovine serum albumin (acBSA, 25 μg/mL)
- Residual LP functional activity in patient plasma measured by C3 deposition ELISA
- Coating buffer: 15 mM Na2CO3, 35 mM NaHCO3, pH 9.6
- Blocking buffer: 1% BSA-TBS (0.1% (w/v) BSA in 10 mM Tris–CL, 140 mM NaCl, 1.5 mM NaN3, pH 7.4)
- Washing buffer: TBS with 0.05% Tween 20 and 5 mM CaCl2
- Barbital buffered saline (BBS): 4 mM barbital, 145 mM NaCl, 2 mM CaCl2, 1 mM MgCl2, pH 7.4
- Incubation time: 1 h 30 min at 37°C for plasma samples, 1 h 30 min at RT for antibodies
- Negative control: Wells receiving only BBS buffer
Annotations
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