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Title: A Research Design for the Quantification of the Neuropeptides Substance P and Calcitonin Gene-Related Peptide in Rat Skin Using Western Blot Analysis
Authors: LAPIN, Guilherme Abbud FrancoHOCHMAN, BernardoNISHIOKA, Michele AkemiMAXIMINO, Jessica RuivoCHADI, GersonFERREIRA, Lydia Masako
Citation: ADVANCES IN SKIN & WOUND CARE, v.28, n.6, p.259-265, 2015
Abstract: OBJECTIVE: To describe and standardize a protocol that overcomes the technical limitations of Western blot (WB) analysis in the quantification of the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) following nociceptive stimuli in rat skin. DESIGN: Male Wistar rats (Rattus norvegicus albinus) weighing 250 to 350 g were used in this study. Elements of WB analysis were adapted by using specific manipulation of samples, repeated cycles of freezing and thawing, more thorough maceration, and a more potent homogenizer; increasing lytic reagents; promoting greater inhibition of protease activity; and using polyvinylidene fluoride membranes as transfer means for skin-specific protein. Other changes were also made to adapt the WB analysis to a rat model. SETTING: University research center. MAIN OUTCOME MEASURE: Western blot analysis adapted to a rat model. RESULTS: This research design has proven effective in collecting and preparing skin samples to quantify SP and CGRP using WB analysis in rat skin. CONCLUSION: This study described a research design that uses WB analysis as a reproducible, technically accessible, and cost-effective method for the quantification of SP and CGRP in rat skin that overcomes technical biases.
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Artigos e Materiais de Revistas Científicas - FM/MNE
Departamento de Neurologia - FM/MNE

Artigos e Materiais de Revistas Científicas - LIM/45
LIM/45 - Laboratório de Fisiopatologia Neurocirúrgica

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