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For subclass experiments, cells were incubated with isotype-specific mouse antibodies and subsequently with an Alexa Fluor 568 anti-mouse IgG.
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These probes may find potential applications in large-scale proteomic experiments whereby subclasses of proteins may be selectively identified.
The antibody subclass we used for this experiment, mouse IgG2a, is a potent inducer of inflammation due to its high affinity for activating receptors [ 5].
Thus, the motorcycle DB makes it possible to predict enzyme reactions based on the class and subclass of metabolic reactions evidenced by experiments mentioned in scientific literature.
To further investigate the physical interaction between enzyme and substrate, the binding of EndoS and EndoS E235Q) to immobilized polyclonal IgG and IgG1 4 subclasses was studied using slot-binding experiments with immobilized IgG probed with EndoS and EndoS E235Q).
This analysis is likely to uncover only very large effects with the present experiment given the smaller subclass sizes involved and the need for stricter testing thresholds to account for the increased number of multiple tests.
According to the large spectrum of different antimicrobial agents [ 29], the class of β-lactamases was represented in the experiments by aminopenicillin, isoxazolylpenicillin and cephalosporin subclasses.
Out of them, 85 from five classical subclasses have been tested with morpholino (MO) knockdown experiments and 57 of them were found to be important in early development of zebrafish.
The ANOVA test was performed to compare CYYR1 expression levels (for CAG- isoform, CAG+ isoform and CAG-/CAG+ ratio) among each different hystological subclass of the tumors studied in the first experiment.
Our preliminary experiments indicate that polyclonal human anti-PCs of the IgM subclass can inhibit uptake of oxLDL in macrophages and also inhibit pro-inflammatory effects mediated by PAF-like lipids generated during LDL oxidation (Frostegård et al. unpublished observation).
Isotype- and subclass-matched mouse Ig were used as negative controls in all the experiments at a concentration of 100 ng/mL.
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