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Mycoplasmas in cell culture are – among other things – affecting cell growth and cell physiology seriously leading to unsatisfying and less significant results. The increasingly widespread use of more sophisticated and sensitive methods for the detection of mycoplasma contamination in cell cultures has resulted in contamination being detected in numerous cultures. This raises the issue of how to eliminate mycoplasma contamination. Naturally, the ideal solution is to discard the contaminated cells. However, if the valuable cells that are stored in liquid nitrogen are also contaminated, a solution is required for eliminating the mycoplasmas and preparing a new cell bank, particularly if the cells are unique and the result of extensive work. The most commonly used method for elimination, inactivation, or suppression of mycoplasmas in cell cultures is treatment with distinct antibiotics. In general, antibiotic therapies do not result in long-lasting, successful elimination. Also, the cytotoxic properties of antibiotics can cause undesirable side effects on eukaryotic cells and may facilitate the development of resistant mycoplasma strains.
The new Mycoplasma-EX method for mycoplasma elimination overcomes all of these drawbacks combining a non-antibiotic and a thoroughly adjusted antibiotic treatment resulting in unique features as compared to other methods and products. Mycoplasma-EX is the first biological reagent that eliminates mycoplasmas by directly killing them, and not just by inhibiting their growth. It is the only anti-mycoplasma agent that can be used to clean virus stocks and most eukaryotic cell cultures directly while showing an extremely low cytotoxicity. Mycoplasma-EX has been shown to be effective with only one treatment, destroying mycoplasmas permanently within 2-3 hours using the non-antibiotic Initial Treatment Reagent only. When using in addition the antibiotic Succession Treatment Reagent, the teatment time extends to – depending on the cell type – two to four weeks. Mycoplasma-Ex is applicable for most cell lines (e.g. Vero, BHK21, GBK, ML, Hep2, 293, CRFK, H9, Molt4, MT-4, Jurkat) and primary cells as well as virus stocks (e.g. SHV-1, BHV-1, HSV-1, VSV, SFV, FCV, MEV). Success of mycoplasma elimination has to be determined after the initial and – if required – succession treatment using a sensitive mycoplasma detection kit.
Benefits of Mycoplasma-EX:– highly effective against M. orale, M. hyorhinis, M. hominis, Acholeplasma laidlawii, M. arginini, M. fermentans, and other mycoplasma species usually encountered as contaminants in cell cultures– combined non-antibiotic and antibiotic treatment– significantly reduced treatment time– ease of use– temperature stable, ready-to-use solution– no additional dosing during the treatment necessary– low cytotoxicity: kills mycoplasmas very efficiently – but is safe for cells
Mycoplasma-EX is a sterile, ready-to-use solution, aliquoted per tube for single use. Store the original container at temperatures of2 to 8°C.
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虽然没有具体用过检测这个抗原的抗体，但是如果希望抗体特异性好点，能检测丰度低的蛋白，还是选Abcam吧，Santa有时候全凭人品了，内参抗体什么的还能凑活，其他的比较勉强。
GAPDH
万能内参
主要看你的目标蛋白的分子量多大，“目标蛋白要与内参蛋白的分子量差异大！”
方便检测
核蛋白，有很多种，分子量不同
膜蛋白，有很多很多种，分子量不同
浆蛋白，有成千上万种，分子量不同。
具体实验，具体目的蛋白，具体的内参。
因为内参有好几种，分子量差别也比较大，更换分子量与检测蛋白差距更大的内参就可以避免这个问题
另外也可以先用一抗孵育显色和检测，再用Strip缓冲液洗掉膜上的抗体，重新进行内参的抗体孵育显色检测。这样也可以将检测蛋白和内参显色在同一张膜上
内参抗体：
Beta-Actin mAb (1C7) 细胞总蛋白 42 kD Abbkine A01010 小鼠源单克隆
Beta-Actin mAb (1C7) , HRP 细胞总蛋白 42 kD Abbkine A01015 小鼠源单克隆,HRP偶联
GAPDH mAb (2B5) 细胞总蛋白 36 kD Abbkine A01020 小鼠源单克隆
GAPDH mAb (2B5) , HRP 细胞总蛋白 36 kD Abbkine A01025 小鼠源单克隆,HRP偶联
Beta-Tubuline mAb (3G6) 细胞总蛋白 55 kD Abbkine A01030 小鼠源单克隆
Plant Actin mAb (3T3) 植物细胞总蛋白 42 kD Abbkine A01050 小鼠源单克隆
PCNA mAb (1D7) 细胞核蛋白 28 kD Abbkine A01040 小鼠源单克隆
COX IV mAb (14Y2) 细胞线粒体总蛋白 16 kD Abbkine A01060 小鼠源单克隆
Histone H3 mAb (2D10) 细胞核蛋白 18 kD Abbkine A01070 小鼠源单克隆
标签抗体（Tag Antibody）可用于检测各种商品化表达载体上的标签序列（如：Myc、Flag、His、GST、HA等），籍以分析目的蛋白的表达含量及其功能；
标签抗体：
Anti-Biotin Antibodies
Anti-Dye Antibodies
Anti-FITC Antibodies
Anti-Fluorescent Protein Antibodies
Anti-HRP Antibodies
Beta Galatosidase Antibodies
FLAG Tag Antibodies
GST Tag Antibodies
HA Tag Antibodies
His Tag Antibodies
Myc Tag Antibodies
TAP Tag Antibodies
V5 Tag Antibodies
【GAPDH】或G3PDH是甘油醛-3-磷酸脱氢酶（ glyceraldehyde-3-phosphate dehydrogenase ）的英文缩写。GAPDH是参与糖酵解的一种关键酶，由4个30-40kDa的亚基组成，分子量146kDa，检测条带大约在36kDa。GAPDH基因几乎在所有组织中都高水平表达，且GAPDH 作为管家基因在同种细胞或者组织中的蛋白质表达量一般是恒定的。
【Western Blot】蛋白质印迹法（免疫印迹试验）。它是分子生物学、生物化学和免疫遗传学中常用的一种实验方法。其基本原理是通过特异性抗体对凝胶电泳处理过的细胞或生物组织样品进行着色。通过分析着色的位置和着色深度获得特定蛋白质在所分析的细胞或组织中表达情况的信息。
β-Actin作为内参是得到了公认的，这是针对大多数组织和细胞来说的，它广泛分布于细胞浆内，表达量非常丰富。Beta-actin由375个氨基酸组成，分子量大小为42-43kDa左右。
β-actin的蛋白水平通常不会发生改变，因此被广泛用于Western时上样量是否一致的参照，也常被用于免疫染色观察细胞的微丝结构。在用作Western的参照时，Actin抗体和Tubulin抗体的主要不同之处在于两者所识别蛋白的分子量不同，这样可以选择合适的参照在同一块胶同一张膜上实现同时检测目标蛋白和参照蛋白。向左转|向右转
