C5a and C5b are protein fragments released from the cleavage of complement component 5 by the C5-convertase protease. C5a is a highly inflammatory peptide that is involved in complement activation, attracting of innate immune cells and histamine release. Abnormal activation of inflammatory complement fragments such as C3a, C4a and C5a can lead to inflammatory disease. C5a binds to the C5a receptor to mediate a variety biological activity. Eculizumab is a monoclonal antibody that binds complement protein C5, inhibiting its cleavage into C5a and C5b. Eculizumab is used to reduce the risk of hemolysis and paroxysmal nocturnal hemoglobinuria.
Product Features
Additional Notes
Strip plates and additional reagents allow for use in multiple experiments
Quantitative protein detection
Establishes normal range
The best products for confirmation of antibody array data
Standard Curves
Spiking & Recovery Results
Linearity Results
Application Notes
Kit Components
Pre-Coated 96-well Strip Microplate
Wash Buffer
Stop Solution
Assay Diluent(s)
Lyophilized Standard
Biotinylated Detection Antibody
Streptavidin-Conjugated HRP
TMB One-Step Substrate
Other Materials Required
Distilled or deionized water
Precision pipettes to deliver 2 µl to 1 µl volumes
Adjustable 1-25 µl pipettes for reagent preparation
100 µl and 1 liter graduated cylinders
Tubes to prepare standard and sample dilutions
Absorbent paper
Microplate reader capable of measuring absorbance at 450nm
Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
Prepare all reagents, samples and standards as instructed in the manual.
Add 100 µl of standard or sample to each well.
Incubate 2.5 h at RT or O/N at 4°C.
Add 100 µl of prepared biotin antibody to each well.
Incubate 1 h at RT.
Add 100 µl of prepared Streptavidin solution to each well.
Incubate 45 min at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Kang S., Tsolmon B., Mann A., et al. Safety evaluation of intravenously administered mono-thioated aptamer against E-selectin in mice. Toxicol Appl Pharmacol. 2015 Aug 15;287(1):86-92. doi: 10.1016/j.taap.2015.05.011
Species:
Mouse
Sample Type:
Plasma (ICR mice treated with anti-eselectin aptamer)
