蛋白质印迹(WB)用于测量不同条件下特定蛋白的变化。在蛋白质印迹(WB)过程中涉及许多步骤,包括样品制备、电泳、蛋白转移、抗体孵育和信号检测。为了解释蛋白质印迹实验的结果,一个贯穿全过程的内参是必须的。
内参可保证每个泳道中加入了相同量的蛋白样品,不同泳道上的蛋白以相同的效率从凝胶转到膜上,不同泳道上抗体孵育(对于一抗而言,必要的话也包括二抗)、信号检测都是一致的。
内参的信号通常用来对目标蛋白的信号进行标准化。需要强调的是,对照蛋白抗体应该与实验抗体在相同的印迹上进行。
不同的样品制备需要不同的内参。下面总结了针对不同样品制备常用的内参,包括全细胞/胞浆蛋白、线粒体、核蛋白、植物组织和血清样本。
针对不同样品制备常用的内参以及它们的分子量
蛋白质印迹(Western Blot)实验中产品推荐:
实验步骤 | 推荐指数 | 货号 | 产品名称 | 规格 |
样本制备 | abs9228 | RIPA (中) | 100ml | |
abs9229 | RIPA (强) | 100ml | ||
abs9230 | RIPA (弱) | 100ml | ||
abs9231 | RIPA (强,无抑制剂) | 100ml | ||
abs9160 | NP-40 | 100ml | ||
abs9195 | 1M Tris-HCl,pH6.8 | 1L | ||
abs9201 | 1M Tris-HCl,pH8.8(更多pH请看爱必信网站) | 1L | ||
abs9117 | SDS | 100ml | ||
★★ | abs9116 | WB/IP 细胞裂解液 | 100ml | |
★★ | abs9239 | WB/IP 细胞裂解液(无抑制剂) | 100ml | |
★★★★ | abs9101 | 红细胞裂解液 | 100ml,250ml,500ml | |
abs812852 | PMSF | 50mg,100mg,200mg | ||
abs816442 | Aprotinin 抑肽酶 | 10mg,25mg,50mg,100mg | ||
abs817902 | Peptatin 胃蛋白酶抑制剂A | 10mg,25mg,50mg,100mg | ||
abs9133 | EDTA | 100ml | ||
abs42017684 | EGTA | 1g,10g | ||
abs817431 | 原钒酸盐 | 100mg,500mg | ||
abs47014865 | β-甘油磷酸盐 | 10g,25g | ||
★★ | abs9161 | 广谱型蛋白酶抑制剂混合物 | 1ml | |
★★ | abs9162 | 广谱型磷酸酶抑制剂混合物 | 1ml | |
蛋白定量 | ★★ | abs9232 | BCA蛋白定量试剂盒 | 500次,2500次 |
★★ | abs9233 | 增强型BCA 蛋白定量试剂盒 | 500次 | |
上样与电泳 | abs9226 | DTT溶液(1mol/L) | 5ml,10ml | |
abs9237 | 2X Laemmli样品缓冲液 | 5ml*2 | ||
abs9238 | 10%SDS十二烷基硫酸钠溶液 | 500ml | ||
★★ | abs953 | 2*上样缓冲液 (适用于SDS-PAGE) | 10ml | |
★★ | abs9301 | 预制胶 10%,10 wells | 5片/盒 | |
★★ | abs9308 | 预制胶 10%,15 wells | 5片/盒 | |
★★ | abs9302 | 预制胶 12%,10 wells | 5片/盒 | |
★★ | abs9309 | 预制胶 12%,15 wells | 5片/盒 | |
abs9303 | 预制胶 15% ,10 wells | 5片/盒 | ||
abs9310 | 预制胶 15% ,15 wells | 5片/盒 | ||
abs9304 | 预制胶 4-15%, 10 wells | 5片/盒 | ||
abs9311 | 预制胶 4-15%, 15 wells | 5片/盒 | ||
★★ | abs9305 | 预制胶 4-20%, 10 wells | 5片/盒 | |
★★ | abs9312 | 预制胶 4-20%, 15 wells | 5片/盒 | |
abs9307 | 预制胶 8%, 15 wells | 5片/盒 | ||
abs9306 | 预制胶 8-20%, 10 wells | 5片/盒 | ||
abs9313 | 预制胶 8-20%, 15 wells | 5片/盒 | ||
NEW | abs9328 | 3-5kd小分子量预制胶,16.5%,10wells | 10片/盒 | |
NEW | abs9329 | 3-5kd小分子量预制胶,16.5%,15wells | 10片/盒 | |
★★★ | abs925 | 蛋白电泳套装 | 盒 | |
abs951 | running buffer | 100ml,1L | ||
★★★★★ | abs923 | 预染蛋白marker, 10-245 KDa | 500ul,5*500ul | |
★★★★★ | abs924 | 预染蛋白marker, 10-180 kDa | 250ul,2*250ul | |
abs966 | Genshare考马斯亮蓝蛋白胶快速染色液 | 500ml | ||
★★★ | abs964 | 考马斯亮蓝染色试剂盒(常规型)R250 | 1kit | |
abs965 | 考马斯亮蓝染色试剂盒(增强型)G250 | 1kit | ||
转膜 | abs931 | PVDF膜0.2 μm | 20张 | |
★★ | abs932 | PVDF膜0.45 μm | 20张 | |
★★ | abs959 | NC膜0.2 μm | 20张 | |
abs960 | NC膜0.45 μm | 20张 | ||
abs950 | 10*电转液 | 100ml,1L | ||
封闭 | abs9174 | 脱脂奶粉 | 100g,500g | |
★★★★★ | abs9155 | BSA | 50g,100g | |
一抗孵育 | ★★ | abs830031 | Beta actin antibody | 50ul,100ul |
abs132001 | Beta actin antibody | 50ug,100ug | ||
abs119680 | Beta Actin Polyclonal Antibody | 50ul,100ul | ||
abs132184 | Beta-Actin antibody | 50ug,100ug | ||
abs137975 | Beta-Actin Mouse Monoclonal Antibody | 50ug,100ug | ||
★★ | abs830030 | GAPDH antibody | 50ul,100ul | |
abs132004 | GAPDH antibody | 50ug,100ug | ||
abs137959 | GAPDH Mouse Monoclonal Antibody | 50ug,100ug | ||
abs118936 | GAPDH Polyclonal Antibody | 50ul,100ul | ||
abs131993 | Tubulin alpha Antibody | 50ug,100ug | ||
★★ | abs830032 | Tubulin Beta antibody | 50ul,100ul | |
abs131994 | Tubulin beta Antibody | 50ug,100ug | ||
abs125209 | Tubulin Beta Polyclonal Antibody | 50ul,100ul | ||
二抗孵育 | ★★ | abs928 | Goat anti-mouse IgG-HRP | 100ul,500ul,1ml |
★★ | abs20002 | Goat anti-rabbit IgG-HRP | 100ul,500ul,1ml | |
★★ | abs20003 | Goat anti-Mouse IgG-FITC | 1ml,500ul,100ul | |
★★ | abs20004 | Goat anti-Rabbit IgG-FITC | 100ul,500ul,1ml | |
★★ | abs20005 | Rabbit anti-Goat IgG-HRP | 100ul,500ul,1ml | |
一抗孵育 | abs830001 | Anti- GST | 50ul,100ul | |
abs830002 | Anti- His Tag抗体 | 50ul,100ul | ||
abs830003 | Anti- His(C-terminal Specific) 抗体 | 50ul,100ul | ||
abs830004 | Anti- Myc 抗体 | 50ul,100ul | ||
abs830005 | Anti- DYKDDDDK | 50ul,100ul | ||
abs830006 | Anti- HA 抗体 | 50ul,100ul | ||
abs830008 | Anti-V5,mAb | 50ul,100ul | ||
abs830010 | Anti- GST,HRP | 50ul | ||
abs830011 | Anti- His Tag (HRP) 抗体 | 50ul | ||
abs830012 | Anti- His(C-terminal Specific) (HRP) 抗体 | 50ul | ||
abs830013 | Anti- Myc (HRP) 抗体 | 50ul | ||
abs830014 | Anti- DYKDDDDK,HRP | 50ul | ||
abs830015 | Anti- HA (HRP) 抗体 | 50ul | ||
abs830016 | Anti- V5 (HRP) 抗体 | 50ul | ||
abs137956 | C-Myc-Tag Mouse Monoclonal Antibody | 50ug,100ug | ||
abs137958 | Flag-Tag Mouse Monoclonal Antibody | 50ug,100ug | ||
显色 | ★★★★★ | abs920 | ECL化学发光检测试剂盒 | 2*25ml,2*50ml,2*250ml |
abs9212 | DAB 显色试剂盒(棕黄色,WB) | 1kit | ||
abs9213 | DAB 显色试剂盒(紫蓝色,WB) | 1kit | ||
其他辅助试剂 | abs952 | 10*TBST | 100ml,1L | |
abs9152 | Tween-20 | 500ml | ||
★★ | abs954 | WB一抗二抗稀释液 | 100ml | |
abs969 | Western一抗二抗去除液(中性) | 250ml | ||
abs968 | Western一抗二抗去除液(弱碱性) | 250ml | ||
abs967 | Western一抗二抗去除液(强碱性) | 250ml | ||
新手* | ★★★★★ | abs926 | WB solution base kit(rabbit) | 1kit |
★★★★★ | abs927 | WB solution base kit(mouse) | 1kit | |
★★★★★ | abs928 | WB solution base kit(goat) | 1kit |
Absin特色产品线: WB相关:ECL发光液、预染marker、预制胶;IHC相关:二抗试剂盒、组化笔;IP/CoIP试剂盒;激动剂/抑制剂;血清、BSA、蛋白酶K、CTB、TTX、CEE;凋亡试剂盒;呼吸爆发试剂盒;ELISA试剂盒;重组蛋白;抗体: 二抗、标签抗体、对照抗体;定制服务(抗体/多肽/蛋白/标记/检测)... |
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