A Research Design for the Quantification of the Neuropeptides Substance P and Calcitonin Gene-Related Peptide in Rat Skin Using Western Blot Analysis

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dc.contributor.authorLapin, Guilherme Abbud Franco [UNIFESP]
dc.contributor.authorHochman, Bernardo [UNIFESP]
dc.contributor.authorNishioka, Michele Akemi [UNIFESP]
dc.contributor.authorMaximino, Jessica Ruivo [UNIFESP]
dc.contributor.authorChadi, Gerson [UNIFESP]
dc.contributor.authorFerreira, Lydia Masako [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionIB CNPq Brazilian Natl Council Sci & Technol Dev
dc.date.accessioned2018-06-15T12:47:34Z
dc.date.available2018-06-15T12:47:34Z
dc.date.issued2015-06-01
dc.description.abstractOBJECTIVE: 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.en
dc.description.affiliationUniv Fed Sao Paulo UNIFESP, Div Plast Surg, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Div Plast Surg, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Dept Expt Neurol, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Sao Paulo, Neurosurg Dept, Lab Pathophysiol LIM45, Sao Paulo, Brazil
dc.description.affiliationIB CNPq Brazilian Natl Council Sci & Technol Dev, Brasilia, DF, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo UNIFESP, Div Plast Surg, Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Div Plast Surg, Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Dept Expt Neurol, Sao Paulo, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Neurosurg Dept, Lab Pathophysiol LIM45, Sao Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent259-265
dc.identifierhttps://dx.doi.org/10.1097/01.ASW.0000465373.42350.c1
dc.identifier.citationAdvances In Skin & Wound Care. Philadelphia: Lippincott Williams & Wilkins, v. 28, n. 6, p. 259-265, 2015.
dc.identifier.doi10.1097/01.ASW.0000465373.42350.c1
dc.identifier.issn1527-7941
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/42125
dc.identifier.wosWOS:000354885100004
dc.language.isoeng
dc.publisherLippincott Williams & Wilkins
dc.relation.ispartofAdvances In Skin & Wound Care
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectNeuropeptidesen
dc.subjectSubstance Pen
dc.subjectCalcitonin gene-related peptideen
dc.subjectWestern blot analysisen
dc.titleA Research Design for the Quantification of the Neuropeptides Substance P and Calcitonin Gene-Related Peptide in Rat Skin Using Western Blot Analysisen
dc.typeinfo:eu-repo/semantics/article
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