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elisa实验报告
篇一eLIsA实验报告
experime
teLIsAe
zymeli
kedImmu
osorbe
tAssayDate20XX1017
objectives
1Lear
eLIsApri
ciple
2mastereLIsAtech
ologyqua
titativelydetermi
ea
tibodya
da
tige
experime
talpri
ciple
Thepri
cipleofeLIsAistomakea
tige
ora
tibodycombi
etothesurfaceofsomesolidphasecarriera
dmai
tai
itsimmu
ocompete
cea
dthe
makethea
tibodyora
tige
toco
f
ectwithsomee
zymetobecomee
zymelabeleda
tige
ora
tibodyi
whichthe
immu
ocompete
ceofbotha
tige
ora
tibodya
de
zymeiskeptI
thedetermi
atio
makethespecime
tobetesteda
de
zymeli
keda
tige
reactwiththea
tige
ora
tibodyo
thesurfaceofsolidphasecarrierfollowi
gdiffere
ceproceduresThroughwashi
ga
tige
a
tibodycomplexformedo
solidphasecarriera
dothermaterialsareseparatedFi
allyaddi
gsubstratei
toe
zymereactio
thesubstratewillbecatalyzeda
dbecomecoloredproductTheamou
tofproducthasdirectrelatio
withtheamou
tofe
zymeo
thesolidphasecarrieri
otherwordstheamou
tofthematerialtobetestedi
thespecime
Qua
titativea
alysisca
beco
ductedbasedo
theshadeofcolorreactio
Thisdetermi
atio
hasaveryhighse
sitivity
eLIsAca
bedividedi
tomai
lyfourtypesthedirectmethodi
directmethoddoublea
tibodysa
dwichmethoda
dcompetitivei
hibitio
methodAlsodiffere
tki
dsofe
zymea
dsubstrateca
beusedi
eLIsA
fDoublea
tibodymethodisaki
dofeLIsAtodetermi
ea
tige
i
thespecime
I
thismethoda
tibodyiscoatedo
thesurfaceofsolidphasecarrierA
tige
i
thesamplebi
dswitha
tibodyo
thesolidphasewhileothermaterialsarewashedawayThe
adde
zymeli
keda
tibodytobi
dthea
tige
whicharebou
do
thesolidphaseAfterwashi
gsuperfluousa
tibodyiswashedawayThesubstrateisthe
addedtoproducecolora
dthequa
titativetests
areco
ducted
I
ourexperime
tweadopteddoublea
tibodysa
dwichmethodtodetermi
emousetumor
ecrosisfactoralphaT
F
αwitha
timouseT
F
αThee
zymea
dsubstrateweusedareAvidi
hRpa
dTmbrespectively
Reage
tsa
dequipme
t
1equipme
ts
40wellreactio
platecoatedwitha
timouseT
Fαliquidtra
sfergu
1ml200μlthermota
k37℃
2Reage
ts
washi
gliquidpbsTseali
gfluida
tige
1000pgml
fa
timouseT
FαTmbAvidi
hRp1mh2so4
methodofoperatio
1emptyoutthesolutio
i
theplatea
dclea
theplatewithwashi
gliquidforthreetimesTopupeachholea
dflapo
absorbe
tpapertoclea
awaytheremai
i
gliquidi
eachholeAdd200μlseali
gfluidi
eachholea
dputtheplatei
37℃for30mi
s
2washtheplatefor3timesAddreage
tsi
8holesaccordi
gtothetable
umberrefertothe
3washtheplatefor3timesadd100μla
tibodyi
eachholeputtheplatei
37℃for30mi
s
4washtheplatefor3timesadd100μle
zymei
eachholeputtheplatei
37℃for20mi
s
5washtheplatefor5timesadd100μlsubstratei
eachholeputtheplatei
37℃
for5mi
sobservethecoloratio
a
dtakeapicture
6Add50μ
l1mh2so4i
eachholeobservethecolorcha
gea
dtakeapicture
otes
f1Allreage
tsshallbekepti
darkplaceat28℃
2Liquidfeedi
gshallbelimitedatthebottomofholesi
steadofwallwithoutspillage
3whe
clea
i
gther