SSR的聚丙烯酰胺凝胶电泳分离与银染检测技术实验报告.pdf

SSR的聚丙烯酰胺凝胶电泳分离与银染检测

技术实验报告

英文回答：

Sodiumdodecylsulphate-polyacrylamidegel

electrophoresis(SDS)andsilverstainingtechniques

experimentreport。

Introduction。

Sodiumdodecylsulphate-polyacrylamidegel

electrophoresis(SDS)isatechniqueusedtoseparate

proteinsbasedontheirmolecularweight.Theproteinsare

firstdenaturedbyboilingtheminasolutioncontaining

SDS,adetergentthatdisruptstheproteinsinteractions

andunfoldsit.Thedenaturedproteinsarethenloadedonto

apolyacrylamidegel,whichisaporousmatrixthatactsas

amolecularsieve.Whenanelectriccurrentisappliedto

thegel,theproteinsmigratethroughthegelatdifferent

rates,withsmallerproteinsmovingfasterthanlarger

proteins.

Silverstainingisasensitivetechniqueusedto

visualizeproteinsonapolyacrylamidegel.Theproteins

arefirstfixedtothegelbysoakingitinasolution

containingglutaraldehyde,across-linkingagentthat

preventstheproteinsfromdiffusing.Thegelisthen

incubatedinasolutioncontainingsilvernitrate,which

bindstotheproteins.Thesilverionsarethenreducedto

silvermetal,whichappearsasadarkbrownbandonthegel.

Materialsandmethods。

Proteinsamples。

SDSgel。

Electrophoresisapparatus。

Silverstainingsolution。

Glutaraldehydesolution。

Gloves。

Safetyglasses。

Procedure。

1.LoadtheproteinsamplesontotheSDSgel.

2.Runtheelectrophoresisapparatusaccordingtothe

manufacturersinstructions.

3.Fixthegelinglutaraldehydesolutionfor30

minutes.

4.Rinsethegelwithwaterfor5minutes.

5.Incubatethegelinsilverstainingsolutionfor30

minutes.

6.Rinsethegelwithwaterfor5minutes.

7.Drythegel.

Results