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Fig. 8 | BMC Neuroscience

Fig. 8

From: Generation of a novel monoclonal antibody that recognizes the alpha (α)-amidated isoform of a valine residue

Fig. 8

Quantification of endogenous valine amide peptide-immunoreactive material in tissues. The abundance of valine amide peptide-immunoreactive material was estimated in purified peptide fractions prepared from subtracted brain regions (CB, CX, BS) and neuroendocrine glands (PIT, PNCR), which displayed moderate-to-high PC18C5 mAb immunolabeling (see Figs. 2 and 7) based on the solid-phase RIA developed for the PC18C5 mAb (inset). The plot shows that the highest abundance of Val-CONH2-ir material was found in the pituitary (PIT) (2.34 ± 0.4672 nmol/μg protein; mean ± SEM), followed by the brainstem (BS) (1.09 ± 0. 021 nmol/μg protein; mean ± SEM), the adrenal gland (AD) (0.35 ± 0.052 nmol/μg protein; mean ± SEM), and the cortex (Cx) (0.41 ± 0.06 nmol/μg protein; mean ± SEM). Inset depicts a representative solid-phase RIA for the PC18C5 mAb used for the quantification of valine amide-immunoreactive material in the assays. Tyr-Gly(3)-Val-CONH2 was used as a standard to construct a typical standard curve and was detected at a concentration of 154.2 fmol/well at the IC50 value, whereas metorphamide/adrenorphin (used as a competitive peptide antigen) was detected at a concentration as low as 7.9 fmol/well at the IC50 value (see text for additional details)

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