蛋白质印迹法（免疫印迹试验）即Western Blot是分子生物学、生物化学和免疫遗传学中常用的一种实验方法。其基本原理是将电泳分离后的细胞或组织蛋白质从凝胶转移到固相支持物例如硝化纤维（NC）或聚偏二氟乙烯（PVDF）膜上，然后通过特异性抗体对样品进行着色。根据分析着色的位置和着色深度获得特定蛋白质在所分析的细胞或组织中表达情况的信息。这种方法现在已经广泛应用于基因在蛋白水平的表达研究、抗体活性检测和疾病早期诊断（例如：HIV）等多个方面。
蛋白质印迹法繁琐复杂，每一个步骤都有很多需要注意的操作。只有小心翼翼做好每一步，才能保证最终实验的成功。顺利跑完电泳后，印迹法的第一步是转渍，也是重要的开端。因为每个人的目标蛋白不同，所以没有统一的转渍protocol让大家使用。转渍的方法选择、实验条件和时间是需要通过实际经验来不断摸索和总结的。那么到底怎样才能确定蛋白质被完整转移了呢？小编今儿就给大家讲解下转渍过程中一些小技巧，不但可以检测转渍效率，也可以让大家在实验上少走不必要的弯路。
1蛋白分子量标准的完整转移
一定要使用提前染色的蛋白质标记（Pre-stained Protein Marker）来监测蛋白质样品的转移。虽然样品中的蛋白质是看不见的，但是如果染色很明显的标准被成功转移到膜上的话，意味着大部分的样品也同样被转移了。有时虽然分子量很大的蛋白质标记没有被转移，大家爷不必太担心，因为目标蛋白一般都比最大的标记分子量小很多，应该是被成功转移了的。其实更值得注意的是转渍时间不宜过长，否则分子量很小（小于20kDa）的蛋白有可能已经从膜的孔隙穿过，附着在滤纸上了。还有一点就是，如果你想在转渍的过程中观察标记转移程度的话，这个小动作很容易引入气泡，影响转渍效率。所以小编建议大家使用不是很重要的样品提前做好优化实验，确定转渍条件和时间，再进行真正的转渍。
2转渍后，把SDS-PAGE的凝胶染色
第二个监测转渍效率的方法就是将凝胶染色，例如Coomassie Stain。这个方法简单、便捷，可以清晰地显现出凝胶上的蛋白质含量，从而判断转渍效率。如果凝胶基本上是透明、干净的，那么转渍很成功；如果凝胶上还有很多蛋白质条带，证明需要增加转渍时间。小编在这里提醒一下大家，如果这么做了的话，即使有很多蛋白还在凝胶上，也不能继续转移到膜上了，因为染色是不可逆的。所以还是建议大家用不是很重要的样品提前做好优化实验，确定转渍条件和时间，再进行真正的转渍。
3对膜进行染色
第二条建议虽然好，但毕竟凝胶染色是不可逆的，会影响实验进度和效率。另外一条建议就是将膜染色。有很多kit是可以成功将膜染色，肉眼就可以辨别出蛋白质的位置，而在短短几分钟内又可以完全将染料去除掉，大家可以放心的进行接下来的实验。这样一来，就可以很好的检测到底有多少蛋白质被转移到膜上。这个方法也会让大家在做接下来的实验时更加有自信。
4转渍在两张膜上
上面的方法都是可以鉴定转渍的时间是否太短。如果大家想检测转渍时间是否设定的过长，就可以尝试着放两张膜，然后进行规范的转渍实验。如果转渍条件合适，那么大部分的蛋白质是应该在离凝胶最近的那张膜上；反之，在另外一张膜上也会检测到不少蛋白。如果你的目标蛋白的分子量很小，那尤其需要注意。蛋白质分子量越小，转渍需要的时长越短。如果在一张膜上找不到目标蛋白，那很有可能已经转移到第二张膜上了！
总结来看，小编建议大家要依据自己目标蛋白的特点来提前优化转渍条件。成功转渍也算是开启蛋白质印迹法成功大门的第一把钥匙啦！
图片来自网络，内容由BioEngX原创
バイアグラ и–¬е±ЂгЃ§иІ·гЃ€г‚‹ – г‚їгѓЂгѓ©гѓ•г‚Јгѓ« жµ·е¤–йЂљиІ© г‚·г‚ўгѓЄг‚№йЂљиІ©
eurax without prescription – aczone cost purchase aczone online cheap
You suggested it superbly!
foreign online casinos online usa casino can online casinos ban you for winning
Wonderful information. Thank you!
yamava online casino online casino for real money usa online casino malaysia 100 welcome bonus
Awesome info. Kudos!
casino online luckia free online casino games slots casino online kz
Thanks a lot. A good amount of forum posts!
best online casino game odds best online casino mentor keno online casino
Perfectly spoken really! .
1 2 gaming online casinos free slot machine online casino sugarhouse casino pa online
Wonderful information. Many thanks!
free bonus online casino no deposit best real money online casinos casino online evolution
Incredible many of beneficial info!
online casino jackpot win new online casinos real money casinos online permitidos em portugal
Wonderful advice. Regards.
new casino sites online best casino online canada online casino real money usa no deposit
Nicely put, Kudos.
the best online casino in usa free slot machines casino online slots games borgata casino online login
Nicely put. Thanks!
ne online casinos online casino real money galaxy world casino online
Wonderful content. Kudos.
a-z online casino online casino live casino online dealer job description
You actually mentioned it perfectly!
ideal online casino online slots casinos resorts casino online casino
Fantastic advice. Thanks!
favbet online casino no deposit online casinos uk regulated online casinos
Very good facts, Appreciate it!
lucky luxe online casino casino games online for real money beetle mania deluxe online spielen casino
Amazing forum posts. Cheers!
monopoly casino game online free online slots casinos casino spiele kostenlos online
Regards. Quite a lot of posts!
online casino georgia best online casino welcome bonus no deposit online casino for canadian players
This is nicely put. !
free online casino india casino online canada online casino reactoonz
“You have not fucked, have you, my son? Have you?” His father asks, as he readjusts the cock covered and swelling in his khaki pants. “I thought as much.”
Awesome stuff. Regards.
malaysia online casino free credit for new member online casino real money usa gta online pc casino glitch
oral betamethasone – purchase differin buy generic monobenzone online
Thanks! I enjoy it!
10 top uk online casino’s casino slots online for real money pacific casino online real money
I was surprised He wasn’t angry. He said,” boy you wait here, I am going to the bathroom.” He closed the door. He must sit on the toilet with pants down because I heard him losing the belt but didn’t hear him peeing. After a while, he called my name,” get in here ass sniffing slut.” I opened the door and crawled to the side of daddy. Daddy was still sitting on the toilet. . Just when I was confused why he sat that and did nothing. He stood up and pulled his pants up. I did take a peep of daddy’s huge uncut Middle Eastern dick. Half hard like 7inches already. While he was doing that, he grabbed my hair pushed right into the toilet, and closed the lid. “ smell my ass from there boy when it is still fresh.” He paused and continued, “ Don’t move.” “I think all of us were sprouting wood, today, dad.” He says. “Coach even noticed how hard we all were.”
buy acticin generic – permethrin price buy generic tretinoin
order generic deltasone – order deltasone for sale zovirax us
With thanks! Useful stuff!
caesar casino online promo code [url=https://usacasinomaster.com/#]top 10 online casinos[/url] play online casino with echeck
Good facts. With thanks.
best casino online south africa [url=https://luckyusaplay.com/#]us casinos online[/url] real casino online for real money no deposit
Regards, Quite a lot of stuff!
z online casino [url=https://luckyusaplay.com/#]online casino usa no deposit bonus[/url] b spot online casino
purchase omnicef online – cleocin sale
Amazing lots of beneficial data.
the most trusted online casino no deposit casino bonus usa online casinos online casino al
Thank you. Useful information.
casino online gratis geld casino online no deposit real money casino slots online
Appreciate it! Plenty of advice.
gry casino maszyny online live online casino river casino online
Thanks a lot, Plenty of advice!
online casino roulette software [url=https://usaplayerscasino.com/#]online american casinos[/url] evolution online casino jobs
With thanks! An abundance of posts.
best nj online casinos [url=https://usacasinomaster.com/#]online casino sign up bonus[/url] online roulette casino
Seriously tons of excellent facts.
mgm online casino canada online casino free play altcoin casino online
You actually stated this fantastically!
www free online casino slot games [url=https://luckyusaplay.com/#]new online casino no deposit bonus[/url] pornhub online casino
Amazing many of fantastic facts!
mo online casino play casino online casino royale movie watch online
Fantastic info. Cheers.
pa online casino 2023 new usa online casinos with no deposit bonuses allslot casino online
Kudos, I value it!
pagcor online casino philippines [url=https://usaplayerscasino.com/#]no deposit online casino[/url] free online casino slots australia
Kudos, Quite a lot of knowledge!
live casino games online slots new online casino usa ok new sc casino online
Really all kinds of excellent material.
online casino free tournaments slot [url=https://usacasinomaster.com/#]new online casinos[/url] online casino kansas
Superb stuff, Many thanks.
casino online argentina pago facil online us casinos online skrill casino
Amazing posts. Kudos.
online casino mit deutscher lizenz online casino game online casino foxwoods
Thanks a lot! I appreciate this!
lady luck online casino no deposit bonus [url=https://uscasinoguides.com/#]best online usa casinos[/url] online casino chat
Excellent data. Thanks a lot!
prism online casino instant play top 10 online casino gta online diamond casino membership
Incredible a lot of terrific advice.
online casino s ДЌeskou licenci play casino games online ameristar casino application jobs & careers online
You actually reported this exceptionally well!
four winds online casino & sportsbook – michigan [url=https://usagamblinghub.com/#]casino bonus online[/url] cabaret club online casino spanish
buy trihexyphenidyl – buy emulgel cheap diclofenac gel purchase online
With thanks. Loads of posts!
online casino high roller bonus gambling casino online online casino free 100 pesos philippines
Whoa many of great material!
online no deposit casinos usa online casino free play no deposit malaysia online casino slot
You actually suggested it very well.
avalon78 online casino [url=https://uscasinoguides.com/#]online casino gaming[/url] how do online casino wagering requirements work
This is nicely said. .
foxwoods online casino real money best online casino no deposit sign up bonus states with legal online casino gambling
Thanks a lot. I enjoy it!
gta online casino macbeth whiskey [url=https://usaplayerscasino.com/#]no deposit online casino[/url] ac online casinos
Terrific stuff, With thanks.
mГ©todos de pago para casinos online populares best online casino no deposit sign up bonus myanmar online casino game
You stated that well!
online casino best deal [url=https://usagamblinghub.com/#]casino online games free[/url] australian registered online casino
Thank you! Good stuff.
online casino technology [url=https://usacasinomaster.com/#]online casinos[/url] bingo online casino philippines
Cheers. Numerous tips!
buy online casino software [url=https://luckyusaplay.com/#]us online casinos with rtg software[/url] online casino roulette cheats
Very well spoken truly. !
3d online casino best online casinos dunder online casino login
Nicely put, Thank you.
seriöses online casino paypal play casino games online best online casino in new zealand
Nicely expressed without a doubt! .
fastest payout online casino ontario best online casino no deposit sign up bonus how to apply online casino dealer
Regards, Numerous stuff!
popular online casino sites newest online casinos usa authorized swiss online casino
Wonderful facts. Thanks a lot.
austria online casinos [url=https://usaplayerscasino.com/#]online casino paypal[/url] mejores casino online chile
Amazing tips. Cheers!
spin city online casino login deposit bonus online casinos nj online casino bonus codes
This is nicely put. .
play’n go online casino [url=https://usacasinomaster.com/#]no deposit casino bonus usa online casinos[/url] online casinos in tennessee
Info clearly applied!!
how to cheat online casinos legit online casinos online casino accepts paypal
Nicely put. Appreciate it.
top online pa casinos [url=https://luckyusaplay.com/#]usa casino online[/url] tropicana casino online app
Nicely put. With thanks!
best new online casinos 2019 new usa online casinos with no deposit bonuses skycity online casino reviews
With thanks. I value it.
online casino bank deposit best online casinos usa online casino slots hack
Thank you, An abundance of forum posts!
uk licensed online casino [url=https://usacasinomaster.com/#]play casino online[/url] online betting and casino
Excellent write ups. Thanks a lot!
free online casino no deposit bonus codes online casino welcome bonus bet mgm online casino phone number
Thanks! An abundance of knowledge.
the casino online [url=https://uscasinoguides.com/#]new online casino no deposit bonus[/url] casinos online que aceptan paysafecard
Superb content. Appreciate it!
best way to win money online casino online casino free bonus no deposit compare uk online casinos
order cyproheptadine 4mg pills – zanaflex drug buy generic zanaflex for sale
You suggested this really well!
online casinos no deposits brand new online casinos usa no deposit bonus codes australian online casino minimum deposit 10
Whoa plenty of valuable tips.
gta online meteorite casino [url=https://usagamblinghub.com/#]online casino gambling[/url] authentic online casino
Cheers. A good amount of info!
online casino roulette canada casino free online casino online 357 play
Valuable information. Many thanks.
using paypal for online casino [url=https://usacasinomaster.com/#]online casino no deposit bonus codes[/url] dragonara casino malta online
Information effectively considered!.
kentucky online casinos online casino with no deposit bonus hard rock casino online real money
You have made your point quite nicely.!
american casino online games [url=https://luckyusaplay.com/#]newest online casinos usa[/url] best online casino no deposit
You have made the point.
trucos para casino online new online usa casinos 918 kiss casino online
Seriously tons of very good material!
real casino slot machines online online casino games free online casino best no deposit bonus
Fantastic content. Kudos!
stake online casino usa [url=https://uscasinoguides.com/#]most trusted online casinos for usa players[/url] raging rhino online casino
Lovely postings, Cheers!
cratos casino online casino free games online playing online casino for real money
Kudos. Very good stuff!
newest online casino pa [url=https://usagamblinghub.com/#]online casino free play[/url] gta online casino masks
You actually mentioned it really well.
play online casino real money australia online casino free play online casino per telefonrechnung
Very good write ups. Cheers!
african online casinos [url=https://luckyusaplay.com/#]casino play online[/url] doubledown casino codes online
Great info. Kudos.
casino online gratis bonus ohne einzahlung new casino online best ar online casino site
Thanks! I value it.
007 casino royale film online https://usaplayerscasino.com/tennis-betting/ 1win online casino canada
mobic 15mg canada – ketorolac usa cheap toradol
Awesome information. With thanks!
online casino sicher gewinnen https://usagamblinghub.com/canadian-casinos/ new casino online no deposit
You explained this fantastically!
novГ© online casino 2023 https://usagamblinghub.com/poker-real-money/ online casino no deposit required usa
You reported that effectively!
play online wind creek casino https://usagamblinghub.com/golf-betting/ online fun casino
You actually revealed that adequately.
online casinos with free play for new members usa https://usaplayerscasino.com/massachusetts-casinos/ online casino blackjack bot
Thanks a lot, Very good information!
take money best online casino india 2022 https://luckyusaplay.com/new-zealand-casinos/ ocean breeze online casino
Thanks a lot! I appreciate it!
m casino online https://usagamblinghub.com/minnesota-casinos/ aprire casino online
buy baclofen 10mg sale – order feldene 20 mg online cheap buy feldene without prescription
Really quite a lot of useful data.
new online casino pa 2021 https://usaplayerscasino.com/betting-apps/ best alabama online casino site
Appreciate it, Plenty of stuff.
casino online bonus de registo https://luckyusaplay.com/ethereum-casinos/ online casino magazine
Position nicely applied.!
maryland online casino signup bonus https://luckyusaplay.com/real-money-casinos/ my choice casino online slots
Awesome forum posts. Appreciate it.
free casino games to play online for fun https://luckyusaplay.com/indiana-casinos/ princess online casino
Superb knowledge. Thanks a lot.
casino online promo codes https://uscasinoguides.com/florida-casinos/ bestes online casino blackjack
voveran tablets – buy nimodipine generic oral nimodipine
You actually said it terrifically!
online casino accept apple pay https://usaplayerscasino.com/real-money-baccarat/ best casino online games free
You’ve made your point pretty clearly!!
online casino mit visa bezahlen https://usagamblinghub.com/superbowl-betting/ australian online casino $10 minimum deposit
This is nicely put. .
online live casinos australia https://usagamblinghub.com/new-casinos/ best legit casino online
Good info. Appreciate it.
mohegan sun online casino pa sign up bonus codes https://usaplayerscasino.com/omaha-poker/ ocean casino online login
You’ve made your position extremely well.!
online casino like luckyland slots https://usagamblinghub.com/north-carolina-casinos/ free online casino table games
pyridostigmine cheap – buy imitrex 50mg generic imuran 25mg oral
You stated that exceptionally well.
https://usagamblinghub.com/legit-casinos/
Well expressed indeed! .
https://uscasinoguides.com/colorado-casinos/
rumalaya medication – brand elavil 10mg order amitriptyline without prescription
yf1it1
duphalac bottles – buy generic mentat for sale buy betahistine 16mg for sale
cyclosporine online – oral methotrexate 5mg colcrys where to buy
buy trileptal 600mg pill – oxcarbazepine 300mg canada buy cheap generic synthroid
purchase speman sale – fincar medication fincar pills
brand terazosin – buy avodart online dapoxetine 30mg pills
order finax for sale – finasteride online buy alfuzosin 10 mg without prescription
buy norfloxacin pill – purchase confido online cheap confido for sale
cheap lasuna pills – buy generic himcolin over the counter brand himcolin
buy atorvastatin no prescription – buy zestril pill buy bystolic 5mg online cheap
purchase atenolol pills – buy betapace 40mg generic coreg 6.25mg usa
buy arava 20mg pills – oral calcium carbonate order cartidin without prescription
cheap rogaine sale – buy finpecia without a prescription purchase finasteride sale
purchase durex gel sale – durex condoms where to buy buy zovirax sale
ondansetron order – ondansetron 8mg cheap oral ropinirole 1mg
buy ascorbic acid for sale – buy bromhexine sale buy prochlorperazine generic
order flexeril pill – brand primaquine enalapril 5mg canada
buy generic cytoxan – antivert 25mg pills vastarel tablets
spironolactone 100mg generic – epitol order online order revia 50mg sale
generic disopyramide phosphate – lyrica over the counter cost thorazine 50 mg
depakote online – cheap generic aggrenox buy cheap generic topamax
piroxicam order online – buy feldene 20 mg for sale buy exelon 3mg for sale
order monograph 600mg generic – monograph cost order cilostazol for sale
buy enalapril 5mg for sale – buy zovirax generic buy zovirax sale
pill dramamine – prasugrel usa pill actonel
order hydroquinone online – order zovirax generic order duphaston 10mg
order dulcolax 5mg online cheap – where can i buy imodium liv52 20mg usa
florinef pills plant – prevacid pills official prevacid pills forehead
promethazine greater – promethazine moon promethazine ponder
ascorbic acid stray – ascorbic acid lazy ascorbic acid ghastly
claritin leader – loratadine chicken claritin tower
priligy travel – priligy batter dapoxetine regular
valtrex online away – valtrex unable valtrex note
uti antibiotics camera – uti treatment minute uti treatment acknowledge
pills for treat prostatitis poor – prostatitis treatment squint prostatitis medications tool
acne medication saturday – acne medication camera acne medication pilot
cenforce online still – levitra professional online pant brand viagra pills study
dapoxetine government – aurogra crack cialis with dapoxetine suck
cialis soft tabs collect – caverta online later viagra oral jelly strong
brand cialis cauldron – viagra soft tabs next penisole dwarf
cenforce performance – cenforce online annoy brand viagra pills shutter
crestor echo – pravachol remain caduet majesty
zocor mister – fenofibrate roger atorvastatin week
nitroglycerin for sale online – nitroglycerin without prescription buy valsartan 80mg online cheap
buy microzide tablets – cheap felodipine buy bisoprolol 10mg without prescription
lopressor 100mg generic – buy generic hyzaar nifedipine 10mg for sale
how to get lanoxin without a prescription – buy cheap calan furosemide over the counter
buy generic nizoral 200mg – buy ketoconazole 200 mg generic order itraconazole pill
buy terbinafine 250mg sale – buy griseofulvin online buy generic griseofulvin
buy generic glyburide for sale – cost dapagliflozin 10mg dapagliflozin online buy
desloratadine canada – cost clarinex albuterol buy online
buy medrol 4 mg – buy loratadine 10mg buy azelastine sprayer
ventolin inhalator cheap – seroflo inhalator online order buy theo-24 Cr
buy azithromycin online cheap – cheap tindamax 300mg oral ciprofloxacin 500mg
amoxil without prescription – axetil canada order baycip online
augmentin pill – buy septra generic buy cheap generic ciprofloxacin
buy atarax generic – escitalopram 10mg canada endep buy online
order quetiapine pill – buy seroquel paypal cheap eskalith pills
order clomipramine 25mg – buy cymbalta 20mg pill buy doxepin 75mg generic
retrovir brand – how to get epivir without a prescription allopurinol 100mg pill
purchase clozaril online – buy clozapine online cheap buy famotidine without a prescription
brand metformin 500mg – buy ciprofloxacin generic lincocin 500mg brand
lasix pills – buy atacand online buy capoten 25mg
order metronidazole 400mg pill – buy oxytetracycline 250mg pills zithromax drug
cost acillin cheap amoxil generic cheap generic amoxil
order valacyclovir pills – nemasole pill acyclovir 400mg us
ivermectin 3 mg for humans – tetracycline us buy sumycin 500mg generic
flagyl online order – buy generic cefaclor 250mg zithromax 500mg pill
order ciprofloxacin 500mg pills – order generic amoxicillin 250mg
order erythromycin 250mg sale
oral ciprofloxacin – how to get myambutol without a prescription buy clavulanate pills
buy baycip pill – order bactrim 480mg without prescription generic augmentin 1000mg
buy atorvastatin sale lipitor 20mg oral order atorvastatin 20mg online