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Alexa Fluor® 555和 Alexa Fluor® 568標(biāo)記二抗——Jackson ImmunoResearch
在進(jìn)行多個(gè)靶標(biāo)的免疫熒光實(shí)驗(yàn)時(shí)，可區(qū)分開的不同光譜的熒光染料和與其他物種交叉吸附的二抗對(duì)于特異性檢測(cè)每個(gè)靶標(biāo)至關(guān)重要。Jackson ImmunoReseach最新推出兩種Alexa Fluor® 標(biāo)記二抗， Alexa Fluor® 555和Alexa Fluor® 568，這兩種染料明亮且易于使用，補(bǔ)充了Jackson現(xiàn)有的Alexa Fluor® 標(biāo)記二抗系列產(chǎn)品線，為橙色通道提供了兩種新的熒光選擇。除了廣譜范圍的熒光外，Jackson ImmunoReseach Alexa Fluor®偶聯(lián)的二抗還具有廣泛的靶特異性。它們與對(duì)許多常見物種交叉反應(yīng)最小的驢或山羊宿主來(lái)源的二抗偶聯(lián)，以避免多種一抗之間的交叉反應(yīng)。
》實(shí)驗(yàn)示例1
DAPI、Alexa Fluor® 488、Alexa Fluor® 555和Alexa Fluor® 647的四色免疫熒光
● 通過(guò)對(duì)人喉表皮樣癌細(xì)胞HEp-2進(jìn)行細(xì)胞內(nèi)蛋白標(biāo)記物染色來(lái)證明該四色熒光組合的可行性（圖1）。
● 使用抗Ki-67（細(xì)胞增殖標(biāo)志物，ref 1），再用Alexa Fluor® 488偶聯(lián)的驢抗兔（H+L）(711-545-152)，綠色顯示細(xì)胞增殖；使用抗α微管蛋白（Mouse Anti-α tubulin）對(duì)微管進(jìn)行染色，再用Alexa Fluor® 555偶聯(lián)的驢抗鼠IgG (H+L) (715-565-150)，觀察到的微管為黃色。順式高爾基體基質(zhì)蛋白參與高爾基體胞室的堆疊和高爾基體結(jié)構(gòu)的維持（ref 2），使用抗人GM130/GOLGA2染色，搭配Alexa Fluor® 647標(biāo)記的驢抗羊IgG (H+L) (713-605-147)，可在細(xì)胞核邊緣看到紅色的順式高爾基體基質(zhì)蛋白。細(xì)胞核用DAPI染色后呈藍(lán)色（圖 2）。
圖 1：DAPI、Alexa Fluor® 488、Alexa Fluor® 555和Alexa Fluor® 647的光譜特性
圖2：人HEp-2細(xì)胞的間接四色免疫染色
》實(shí)驗(yàn)示例2
DAPI、Alexa Fluor® 488、Alexa Fluor® 568、Alexa Fluor® 647的四色免疫熒光
● 通過(guò)對(duì)人喉表皮樣癌細(xì)胞HEp-2進(jìn)行細(xì)胞內(nèi)蛋白標(biāo)記物染色來(lái)證明該四色熒光組合的可行性（圖3）。
● 使用抗Ki-67（細(xì)胞增殖標(biāo)志物，ref 1），再用Alexa Fluor® 488偶聯(lián)的驢抗兔（H+L）(711-545-152)，綠色顯示細(xì)胞增殖；使用抗α微管蛋白（Mouse Anti-α tubulin）對(duì)微管進(jìn)行染色，然后用Alexa Fluor® 568標(biāo)記的驢抗鼠IgG (H+L) (715-575-151)，微管呈黃色。使用上皮細(xì)胞標(biāo)記物E-Cadherin抗體，然后用Alexa Fluor® 647偶聯(lián)的驢抗山羊 IgG (H+L)
● 抗體(705-605-147)，以紅色顯示細(xì)胞間接觸。細(xì)胞核用DAPI染色，可見藍(lán)色（圖4）。
圖 3：DAPI, Alexa Fluor® 488, Alexa Fluor® 568, Alexa Fluor® 647的光譜特性
圖4：人HEp-2細(xì)胞的間接四色免疫染色
》部分產(chǎn)品列表
● Alexa Fluor® 555標(biāo)記二抗
產(chǎn)品貨號(hào) | 產(chǎn)品名稱 | 規(guī)格 |
111-565-144 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Rabbit IgG (H+L) (minimal cross-reaction to Human, Mouse, and Rat Serum Proteins) | 1mg |
112-565-071 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Rat IgG, Fcγ Fragment Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
112-565-075 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Rat IgM, µ Chain Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
112-565-143 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Rat IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, and Rabbit Serum Proteins) | 1mg |
112-565-167 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Rat IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, Mouse, and Rabbit Serum Proteins) | 0.5mg |
115-565-071 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Fragment Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
115-565-075 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgM, µ Chain Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
115-565-146 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, Rabbit, and Swine Serum Proteins) | 1mg |
115-565-166 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, Rabbit, and Rat Serum Proteins) | 0.5mg |
115-565-205 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
115-565-206 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 2a Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
115-565-207 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 2b Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
115-565-209 | Alexa Fluor® 555-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
300-565-245 | Alexa Fluor® 555-conjugated AffiniPure™ Rabbit Anti-GFP | 0.1mg |
609-564-213 | Alexa Fluor® 555-conjugated AffiniPure™ VHH Alpaca Anti-Human IgG (H+L) (minimal cross-reaction to Bovine, Mouse, and Rabbit Serum Proteins) | 0.1mg |
611-564-215 | Alexa Fluor® 555-conjugated AffiniPure™ VHH Alpaca Anti-Rabbit IgG (H+L) (minimal cross-reaction to Bovine, Human, and Mouse Serum Proteins) | 0.1mg |
615-564-214 | Alexa Fluor® 555-conjugated AffiniPure™ VHH Alpaca Anti-Mouse IgG (H+L) (minimal cross-reaction to Bovine, Human, and Rabbit Serum Proteins) | 0.1mg |
703-565-155 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Chicken IgY (IgG) (H+L) (minimal cross-reaction to Bovine, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, Rat, and Sheep Serum Proteins) | 0.5mg |
705-565-147 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Goat IgG (H+L) (minimal cross-reaction to Chicken, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins) | 0.5mg |
706-565-148 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Guinea Pig IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins) | 0.5mg |
709-565-149 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Human IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Mouse,Rabbit,Rat,and Sheep Serum Proteins) | 0.5mg |
711-565-152 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Rabbit IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig,Syrian Hamster,Horse, Human,Mouse,Rat,and Sheep Serum Proteins) | 0.5mg |
712-565-150 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Rat IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Rabbit, and Sheep Serum Proteins) | 0.5mg |
712-565-153 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Rat IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Sheep Serum Proteins) | 0.5mg |
713-565-147 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Sheep IgG (H+L) (minimal cross-reaction to Chicken, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins) | 0.5mg |
715-565-150 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Mouse IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Rabbit, and Sheep Serum Proteins) | 0.5mg |
715-565-151 | Alexa Fluor® 555-conjugated AffiniPure™ Donkey Anti-Mouse IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Rabbit, Rat, and Sheep Serum Proteins) | 0.5mg |
● Alexa Fluor® 568標(biāo)記二抗
產(chǎn)品貨號(hào) | 產(chǎn)品名稱 | 規(guī)格 |
111-575-144 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Rabbit IgG (H+L) (minimal cross-reaction to Human, Mouse, and Rat Serum Proteins) | 1mg |
112-575-071 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Rat IgG, Fcγ Fragment Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
112-575-075 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Rat IgM, µ Chain Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
112-575-143 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Rat IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, and Rabbit Serum Proteins) | 1mg |
112-575-167 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Rat IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, Mouse, and Rabbit Serum Proteins) | 0.5mg |
115-575-071 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Fragment Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
115-575-075 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgM, µ Chain Specific (minimal cross-reaction to Human, Bovine, and Horse Serum Proteins) | 1mg |
115-575-146 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, Rabbit, and Swine Serum Proteins) | 1mg |
115-575-166 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG (H+L) (minimal cross-reaction to Human, Bovine, Horse, Rabbit, and Rat Serum Proteins) | 0.5mg |
115-575-205 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
115-575-206 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 2a Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
115-575-207 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 2b Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
115-575-209 | Alexa Fluor® 568-conjugated AffiniPure™ Goat Anti-Mouse IgG, Fcγ Subclass 3 Specific (minimal cross-reaction to Human, Bovine, and Rabbit Serum Proteins) | 0.5mg |
300-575-245 | Alexa Fluor® 568-conjugated AffiniPure™ Rabbit Anti-GFP | 0.1mg |
609-574-213 | Alexa Fluor® 568-conjugated AffiniPure™ VHH Alpaca Anti-Human IgG (H+L) (minimal cross-reaction to Bovine, Mouse, and Rabbit Serum Proteins) | 0.1mg |
611-574-215 | Alexa Fluor® 568-conjugated AffiniPure™ VHH Alpaca Anti-Rabbit IgG (H+L) (minimal cross-reaction to Bovine, Human, and Mouse Serum Proteins) | 0.1mg |
615-574-214 | Alexa Fluor® 568-conjugated AffiniPure™ VHH Alpaca Anti-Mouse IgG (H+L) (minimal cross-reaction to Bovine, Human, and Rabbit Serum Proteins) | 0.1mg |
703-575-155 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Chicken IgY (IgG) (H+L)(minimal cross-reaction to Bovine, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, Rat, and Sheep Serum Proteins) | 0.5mg |
705-575-147 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Goat IgG (H+L) (minimal cross-reaction to Chicken, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins) | 0.5mg |
706-575-148 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Guinea Pig IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins) | 0.5mg |
709-575-149 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Human IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Mouse, Rabbit, Rat, and Sheep Serum Proteins) | 0.5mg |
711-575-152 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Rabbit IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rat, and Sheep Serum Proteins) | 0.5mg |
712-575-150 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Rat IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Rabbit, and Sheep Serum Proteins) | 0.5mg |
712-575-153 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Rat IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Sheep Serum Proteins) | 0.5mg |
713-575-147 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Sheep IgG (H+L) (minimal cross-reaction to Chicken, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, and Rat Serum Proteins) | 0.5mg |
715-575-150 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Mouse IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Rabbit, and Sheep Serum Proteins) | 0.5mg |
715-575-151 | Alexa Fluor® 568-conjugated AffiniPure™ Donkey Anti-Mouse IgG (H+L) (minimal cross-reaction to Bovine, Chicken, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Rabbit, Rat, and Sheep Serum Proteins) | 0.5mg |
》參考文獻(xiàn)
1、Scholzen, T., & Gerdes, J. (2000). The Ki-67 protein: from the known and the unknown. Journal of cellular physiology, 182(3), 311–322. /(SICI)1097-4652(200003)182:3<311::aid-jcp1>3.0.CO;2-9
2、Chang, S. H., Hong, S. H., Jiang, H. L., Minai-Tehrani, A., Yu, K. N., Lee, J. H., Kim, J. E., Shin, J. Y., Kang, B., Park, S., Han, K., Chae, C., & Cho, M. H. (2012). GOLGA2/GM130, Cis-Golgi matrix protein, is a novel target of anticancer gene therapy. Molecular therapy : the journal of the American Society of Gene Therapy, 20(11), 2052–2063.
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