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Alexa Fluor® 555和 Alexa Fluor® 568標記二抗——Jackson ImmunoResearch
在進行多個靶標的免疫熒光實驗時，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于特異性檢測每個靶標至關(guān)重要。Jackson ImmunoReseach最新推出兩種Alexa Fluor® 標記二抗， Alexa Fluor® 555和Alexa Fluor® 568，這兩種染料明亮且易于使用，補充了Jackson現(xiàn)有的Alexa Fluor® 標記二抗系列產(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。它們與對許多常見物種交叉反應(yīng)最小的驢或山羊宿主來源的二抗偶聯(lián)，以避免多種一抗之間的交叉反應(yīng)。
》實驗示例1
DAPI、Alexa Fluor® 488、Alexa Fluor® 555和Alexa Fluor® 647的四色免疫熒光
● 通過對人喉表皮樣癌細胞HEp-2進行細胞內(nèi)蛋白標記物染色來證明該四色熒光組合的可行性（圖1）。
● 使用抗Ki-67（細胞增殖標志物，ref 1），再用Alexa Fluor® 488偶聯(lián)的驢抗兔（H+L）(711-545-152)，綠色顯示細胞增殖；使用抗α微管蛋白（Mouse Anti-α tubulin）對微管進行染色，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標記的驢抗羊IgG (H+L) (713-605-147)，可在細胞核邊緣看到紅色的順式高爾基體基質(zhì)蛋白。細胞核用DAPI染色后呈藍色（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細胞的間接四色免疫染色
》實驗示例2
DAPI、Alexa Fluor® 488、Alexa Fluor® 568、Alexa Fluor® 647的四色免疫熒光
● 通過對人喉表皮樣癌細胞HEp-2進行細胞內(nèi)蛋白標記物染色來證明該四色熒光組合的可行性（圖3）。
● 使用抗Ki-67（細胞增殖標志物，ref 1），再用Alexa Fluor® 488偶聯(lián)的驢抗兔（H+L）(711-545-152)，綠色顯示細胞增殖；使用抗α微管蛋白（Mouse Anti-α tubulin）對微管進行染色，然后用Alexa Fluor® 568標記的驢抗鼠IgG (H+L) (715-575-151)，微管呈黃色。使用上皮細胞標記物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)，以紅色顯示細胞間接觸。細胞核用DAPI染色，可見藍色（圖4）。
圖 3：DAPI, Alexa Fluor® 488, Alexa Fluor® 568, Alexa Fluor® 647的光譜特性
圖4：人HEp-2細胞的間接四色免疫染色
》部分產(chǎn)品列表
● Alexa Fluor® 555標記二抗
產(chǎn)品貨號 | 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標記二抗
產(chǎn)品貨號 | 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
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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