二抗是能和一抗結(jié)合的抗體，即抗體的抗體。主要用于檢測(cè)一抗的存在。一抗特異性結(jié)合抗原的，二抗是特異性結(jié)合一抗。即一抗的Fc段或Fab片段可以充當(dāng)抗原刺激機(jī)體產(chǎn)生抗體。也就是說(shuō)，一抗主要是IgG類免疫球蛋白，因此二抗就是抗IgG抗體。
一抗二抗都是一種可以特異結(jié)合別的物質(zhì)的基團(tuán)，而且一抗可以至少結(jié)合兩種其他基團(tuán)（底物和二抗），如下圖是直標(biāo)一抗。
一抗是特異性的結(jié)合目標(biāo)蛋白，一抗需要發(fā)光物質(zhì)或者顯色基團(tuán)標(biāo)記才能被檢測(cè)出來(lái)，否者一抗和蛋白的結(jié)合與否用肉眼是看不出來(lái)的。但是一抗往往比較特異每個(gè)都標(biāo)記成本很高，通常會(huì)使用標(biāo)記二抗可以和一抗結(jié)合，并帶有可以被檢測(cè)出的標(biāo)記(如帶熒光、放射性、化學(xué)發(fā)光或顯色基團(tuán)），所以如今的設(shè)計(jì)一般是二抗帶上可檢測(cè)標(biāo)記，再來(lái)檢測(cè)一抗，而一抗識(shí)別底物。這樣，當(dāng)一抗結(jié)合到底物上，就可以通過(guò)二抗檢測(cè)出來(lái)。
比如ECL 顯色原理：在 ECL 底物中，含有 H2O2 和魯米諾（及其衍生物），在 HRP（辣根過(guò)氧化物酶）的作用下，發(fā)出熒光。
二抗是由一抗種屬（比如兔，小鼠）的某一亞型的免疫球蛋白IgG免疫哺乳動(dòng)物（一般是免疫山羊、馬、驢、騾和豚鼠及小鼠等動(dòng)物）產(chǎn)生的抗體，因此二抗只結(jié)合特定的物種特定亞型的一抗。比如，羊抗小鼠（二抗）會(huì)特異性結(jié)合小鼠單抗（一抗）而不會(huì)結(jié)合兔多抗或大鼠單抗，所以二抗和一抗要匹配上。
實(shí)驗(yàn)方法不同，二抗耦聯(lián)的基團(tuán)就會(huì)不同，通常主要有酶(辣根過(guò)氧化酶HRP和堿性磷酸酶AP)，熒光基團(tuán)(FITC、 Rhodamine、PE、Dylight、Alexa Fluor 、cy3等)、生物素、金顆粒、。一般做WB、 ELISA、 IHC等實(shí)驗(yàn)都是推薦HRP標(biāo)記的二抗，IHC推薦生物素標(biāo)記的二抗，做IF、 FC等都是熒光如Alexa Fluor 488、FITC、PE標(biāo)記的抗體。
a.免疫印記（WB）
b.免疫組化（IHC/ICC）
c.免疫熒光（IHF/ICF)
d.免疫沉淀免疫共沉淀
特殊二抗
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標(biāo)記基團(tuán) |
產(chǎn)品貨號(hào) |
產(chǎn)品名稱 |
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無(wú)標(biāo)記 |
Goat Anti-Rabbit lgG(H+L) |
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Goat Anti-Mosue lgG(H+L) |
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Donkey Anti-Human IgG(H+L) |
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HRP |
Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) |
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Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) |
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Peroxidase AffiniPure Goat Anti-Rat IgG (H+L) |
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Peroxidase AffiniPure Goat Anti-Guinea Pig IgG(H+L) |
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Horseradish Peroxidase Goat Anti-Human IgG (H+L) |
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Peroxidase AffiniPure Rabbit Anti-Goat IgG (H+L) |
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Peroxidase AffiniPure Rabbit Anti-Sheep IgG (H+L) |
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Peroxidase AffiniPure Donkey Anti-Rabbit IgG (H+L) |
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Peroxidase AffiniPure Donkey Anti-Mouse IgG (H+L) |
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Peroxidase AffiniPure Donkey Anti-Goat IgG (H+L) |
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Biotin |
Biotin-SP AffiniPure Goat Anti-Rabbit IgG (H+L) |
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Biotin-SP AffiniPure Goat Anti-Mouse IgG (H+L) |
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Biotin-SP AffiniPure Goat Anti-Rat IgG (H+L) |
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Biotin-SP AffiniPure Donkey Anti-Goat IgG (H+L) |
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綠色熒光 |
YSFluor™ 488 Goat Anti-Rabbit IgG (H+L) |
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YSFluor™ 488 Goat Anti-Human IgG (H+L) |
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YSFluor™488 Goat Anti-Mouse IgG (H+L) |
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YSFluor™ 488 Goat Anti-Bovine IgG (H+L) |
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YSFluor™ 488 Goat Anti-Guinea Pig IgG (H+L) |
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YSFluor™488 Rabbit Anti-Goat IgG (H+L) |
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YSFluor™ 488 Rabbit Anti-Mouse IgG (H+L) |
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YSFluor™488 Donkey Anti-Sheep IgG (H+L) |
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YSFluor™488 Donkey Anti-Mouse IgG (H+L) |
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YSFluor™488 Donkey Anti-Rabbit IgG (H+L) |
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YSFluor™ 488 Donkey Anti-Goat IgG (H+L) |
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YSFluor™ 488 Donkey Anti-Rat IgG (H+L) |
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YSFluor™488 Donkey Anti-Guinea Pig IgG (H+L) |
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YSFluor™488 Donkey Anti-Chicken IgG (H+L) |
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YSFluor™488 Rabbit Anti-Chicken IgY(H+L) |
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YSFluor™488 Donkey Anti-Human IgG (H+L) |
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FITC AffiniPure Goat Anti-Human IgG(H+L) |
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FITC-AffiniPure Goat Anti-Rabbit IgG (H+L) |
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FITC-AffiniPure Goat Anti-Mouse IgG (H+L) |
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FITC-AffiniPure Goat Anti-Rat IgG (H+L) |
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FITC-AffiniPure Rabbit Anti-Goat IgG (H+L) |
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FITC-AffiniPure Rabbit Anti-Sheep IgG (H+L) |
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羅丹明 |
Rhodamine (TRITC) Goat Anti-Rabbit IgG (H+L) |
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Rhodamine (TRITC) Goat Anti-Mouse IgG (H+L) |
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Rhodamine(TRITC) AffiniPure Rabbit Anti-Goat IgG (H+L) |
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Rhodamine Red-X (RRX) Goat Anti-Mouse IgG (H+L) |
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橙色熒光 |
YSFluor™594 AffiniPure Goat Anti-Rabbit IgG (H+L) |
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YSFluor™ 594 Goat Anti-Mouse IgG (H+L) |
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YSFluor™ 594 Goat Anti-Human IgG (H+L) |
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YSFluor™594 Goat Anti-Guinea Pig IgG (H+L) |
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YSFluor™ 594 Goat Anti-Bovine IgG (H+L) |
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YSFluor™594 Rabbit Anti-Goat IgG (H+L) |
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YSFluor™ 594 Rabbit Anti-Sheep IgG (H+L) |
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YSFluor™594 Rabbit Anti-Mouse IgG (H+L) |
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YSFluor™594 Rabbit Anti-Chicken IgY (H+L) |
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YSFluor™ 594 Donkey Anti-Mouse IgG (H+L) |
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YSFluor™594 Donkey Anti-Rabbit IgG (H+L) |
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YSFluor™ 594 Donkey Anti-Goat IgG (H+L) |
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YSFluor™ 594 Donkey Anti-Rat IgG (H+L) |
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YSFluor™594 Donkey Anti-Guinea Pig IgG (H+L) |
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YSFluor™594 Donkey Anti-Chicken IgG (H+L) |
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YSFluor™594 Donkey Anti-Human IgG (H+L) |
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YSFluor™594 Donkey Anti-Sheep IgG (H+L) |
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Cy3-AffiniPure Goat Anti-Mouse IgG (H+L) |
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Cy3-AffiniPure Goat Anti-Rat IgG (H+L) |
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Cy3-AffiniPure Rabbit Anti-Goat IgG (H+L) |
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Cy3-AffiniPure Rabbit Anti-Sheep IgG (H+L) |
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Cy3-AffiniPure Donkey Anti-Goat IgG(H+L) |
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紅色熒光 |
YSFluor™647 Goat Anti-Rabbit IgG (H+L) |
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YSFluor™ 647 Goat Anti-Mouse IgG (H+L) |
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YSFluor™647 Goat Anti-Human IgG (H+L) |
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YSFluor™ 647 Goat Anti-Bovine IgG (H+L) |
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YSFluor™ 647 Goat Anti-Guinea Pig IgG (H+L) |
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YSFluor™ 647 Rabbit Anti-Goat IgG (H+L) |
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YSFluor™647 Rabbit Anti-Sheep IgG (H+L) |
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YSFluor™ 647 Rabbit Anti-Mouse IgG (H+L) |
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YSFluor™647 Rabbit Anti-Chicken IgY (H+L) |
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YSFluor™ 647 Donkey Anti-Mouse IgG (H+L) |
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YSFluor™ 647 Donkey Anti-Rabbit IgG (H+L) |
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YSFluor™ 647 Donkey Anti-Rat IgG (H+L) |
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YSFluor™647 Donkey Anti-Human IgG (H+L) |
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YSFluor™647 Donkey Anti-Sheep IgG (H+L) |
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