【储存条件】 -20˚C恒温长期保存，4˚C保存1个月，建议分装保存，避免反复冻融。
【产品简介】
AFT Hiper Multi-color Self-Lighting Western Protein Marker (10-200 kDa) is a ready-to-use mixture with 10 IgG-binding proteins covering a wide range of molecular weights from 10 to 200 kDa in Tris Glycine buffer. AFT Hiper Multi-color Self-Lighting Western Protein Marker (10-200 kDa) performs dual functions. First, it contains 11 pre-stained proteins (10, 15, 25, 35, 45, 65, 70, 75, 100, 135 and 180 kDa) for monitoring protein separation during SDS-PAGE, verification of Western transfer efficiency on membranes (nitrocellulose, PVDF, or nylon) and for approximating the protein size. Second, 10 IgG binding proteins can be immuno-detected on film or by CCD imaging. AFT Hiper Multi-color Self-Lighting Western Protein Marker (10-200 kDa) is compatible for chemiluminescent, fluorescent, chromogenic or other detection systems. In addition, AFT Hiper Multi-color Self-Lighting Western Protein Marker (10-200 kDa) has two reference bands with enhanced intensity (at 30 kDa and 80 kDa). The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading.
【使用方法】
1. 将本产品为即可用型产品，在室温融化后，轻柔混匀，使沉淀充分溶解；
2. 加入5 μl到本产品 SDS-聚丙烯酰胺胶的上样孔中，与待测样品一起电泳和转膜；
注意：上样量可根据胶厚度、上样孔宽度、一抗和二抗的种类、滴度以及ECL液的灵敏度等相关因素
进行适当的调整，一般在5-10μl上样，不可低于5μl。
3. 转膜后，进行正常的封闭、一抗和二抗孵育，最后进行ECL发光检测。
4. ECL后，可以同时检测到10个自发光条带和目的条带（如右图所示）。
【注意事项】
1. 使用前先恢复至室温使沉淀充分溶解，否则可能导致电泳条带出现不同程度的弥散或拖带；
2. 本产品含有 SDS，蛋白已变性，不宜作为天然蛋白分子电泳时的分子量参照标准。
【附录：转膜和洗膜】
A、转膜条件（冰上进行）：
a. Transfer with buffer containing 20% methanol to fix proteins on membrane；
b. Wash membrane with PBS or TBS containing less than 0.1% Tween-20 at 4°C。
B、洗膜条件（4度进行）：
Membrane: Nitrocellulose membranes / PVDF；
Wash Buffer:1X Tris buffered saline, 0.1% Tween-20 (TBST)（吐温20不能超过0.1%）。
C、Stripping Buffer:
15g Glycine, 1g SDS, 10 ml Tween20, pH2.2, Adjust volume to 1L。如果需要用到含有DTT /b-ME 的Stripping buffer，膜需要先以 1X Tris buffered saline(TBS) 洗三次，把 Tween-20去干净后，再进行Stripping。 1X Tris buffered saline（6.05 g Tris and 8.76 g NaCl in 800 mL of H2O. Adjust pH to 7.5 with 1 M HCl and make volume up to 1 L with H2O）。
