使用说明：
对于培养细胞样品：
1.融解Western及IP细胞裂解液，混匀。取适当量的裂解液，在使用前数分钟内加入PMSF，使PMSF的最 终浓度为1mM。
2.对于贴壁细胞：去除培养液，用PBS、生理盐水或无血清培养液洗一遍(如果血清中的蛋白没有干扰，可以不洗)。按照6孔板每孔加入100-200微升裂解液的比例加入裂解液。用枪吹打数下，使裂解液和细胞充分接触。通常裂解液接触细胞1-2秒后，细胞就会被裂解。
对于悬浮细胞：离心收集细胞，用手指把细胞用力弹散。按照6孔板每孔细胞加入100-200微升裂解液的比例加入裂解液。再用手指轻弹以充分裂解细胞。充分裂解后应没有明显的细胞沉淀。如果细胞量较多，必需分装成50-100万细胞/管，然后再裂解。大团的细胞较难裂解充分，而少量的细胞由于
裂解液容易和细胞充分接触，相对比较容易裂解充分。
3.充分裂解后，10000-14000g离心3-5分钟，取上清，即可进行后续的PAGE、Western、免疫沉淀和免疫共沉淀等操作。
裂解液用量说明：通常6孔板每孔细胞加入100微升裂解液已经足够，但如果细胞密度非常高可以适当加大裂解液的用量到150微升或200微升。每100万细胞用100微升本产品裂解后获得的上清，其蛋白浓度约为2-4mg/ml，不同细胞有所不同。
对于组织样品：
1.把组织剪切成细小的碎片。
2.融解Western及IP细胞裂解液，混匀。取适当量的裂解液，在使用前数分钟内加入PMSF，使PMSF的最终浓度为1mM。
3.按照每20毫克组织加入100-200微升裂解液的比例加入裂解液。(如果裂解不充分可以适当添加更多的裂解液，如果需要高浓度的蛋白样品，可以适当减少裂解液的用量。)
4.用玻璃匀浆器匀浆，直至充分裂解。
5.充分裂解后，10000-14000g离心3-5分钟，取上清，即可进行后续的PAGE、Western、免疫沉淀和免疫共沉淀等操作。每20mg冻存的小鼠肝脏组织用200微升本裂解液裂解后获得的上清，其蛋白浓度约为15-25mg/ml，不同状态的不同组织有所不同。
6.如果组织样品本身非常细小，可以适当剪切后直接加入裂解液裂解，通过强烈vortex使样品裂解充分。然后同样离心取上清，用于后续实验。直接裂解的优点是比较方便，不必使用匀浆器，缺点是不如使用匀浆器那样裂解得比较充分。

附录：碧云天生产的各种裂解液主要特点、差异和选择
首先请参考下表，了解各种裂解液的主要特点和差异。

    用于普通的Western、IP或co-IP，我们推荐使用Western及IP细胞裂解液(P0013)，该裂解液已被国内各大研究机构广泛使用，发表大量SCI论文，用户普遍反映很好。裂解细胞或组织后，没有非常粘滞的透明状DNA团块形成，不必采用超声处理等就可以非常理想地用于后续操作。另外该裂解液裂解的产物也适合用于磷酸化蛋白的Western检测。
    对于某些特殊蛋白的IP，如果发现Western及IP细胞裂解液(P0013)效果不是非常理想，可以尝试用RIPA裂解液(强、中或弱)或NP-40裂解液。如果发现IP的时候背景很高，即非特异的蛋白也被IP下来，则需要选用裂解强度较高的裂解液，例如RIPA裂解液(强或中)。如果发现目的蛋白无法被IP下来，则说明裂解液的强度过强，可以使用较为温和的裂解液例如RIPA裂解液(弱)或NP-40裂解液。
对于某些难溶解蛋白的Western，如果发现Western及IP细胞裂解液(P0013)效果不是非常理想，可以尝试使用裂解强度更高的裂解液例如RIPA裂解液(强、中)或SDS裂解液。使用RIPA裂解液(强)的用户也非常多，发表了大量SCI论文。
    用于特定用途需要自行添加特定抑制剂或不需要添加抑制剂时，可以考虑选购P0013J或P0013K。P0013J在很多时候可以兼容酶活性和生物小分子的检测，对于特定的酶或生物小分子的检测是否兼容需 要自行测试，碧云天不提供具体的应用信息。P0013J的裂解能力比P0013K弱一些，但用于酶活性和生物小分子时，P0013J的兼容性通常会更好一些。

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77.Yan J, Feng Z, Liu J, Shen W, Wang Y, Wertz K, Weber P, Long J, Liu J.
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84.Zhao Q, Zhang C, Wang X, Chen L, Ji H, Zhang Y.
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99.Zhang R, Shi H, Chen Z, Feng W, Zhang H, Wu K.
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100.Ren J, Zhao J, Zhou Y, Liu X, Chen X, Hu K.
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101.Mao J, Yuan J, Wang L, Zhang H, Jin X, Zhu J, Li H, Xu B, Chen L.
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106.Sun J, Fan L, Li M, Zhang Y, Cheng N.
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107.Qing Y, Liang Y, Du Q, Fan P, Xu H, Xu Y, Shi N.
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108.Li Z, Zhang L, Zhao Y, Li H, Xiao H, Fu R, Zhao C, Wu H, Li Z.
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109. Liu Y, Zhu Z, Xiong Z, Zheng J, Hu Z, Qiu J.
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110.Duan W, Guo Y, Xiao J, Chen X, Li Z, Han H, Li C.
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111.Ge C, Song J, Chen L, Wang L, Chen Y, Liu X, Zhang Y, Zhang L, Zhang M.
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112.Li Z, Wang Y, Wu H, Zhang L, Yang P, Li Z.
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113.Li P, Ji M, Lu F, Zhang J, Li H, Cui T, Li Wang X, Tang D, Ji C.
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115.Zhang JW, Zhang SS, Song JR, Sun K, Zong C, Zhao QD, Liu WT, Li R, Wu MC, Wei LX.
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116.Liu SS, Wang XP, Li XB, Liang JY, Liu LL, Lu Y, Zhong XY, Chen YX.
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117.Zheng A, Li H, Wang X, Feng Z, Xu J, Cao K, Zhou B, Wu J, Liu J.
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118.Zhao L, Zou X, Feng Z, Luo C, Liu J, Li H, Chang L, Wang H, Li Y, Long J, Gao F, Liu J.
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119.Miao CG, Yang YY, He X, Huang C, Huang Y, Qin D, Du CL, Li J.
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120.Ren J, Jiang H, Zhao J, Xin W, Xu Y, Chen X, Hu K.
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121.An P, Ding Y, Wang A, Tan W, Gao F, Fan A, Tang B, Zhang X, Li Z.
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122.Wang C, Pan Y, Zhang R, Bai B, Chen J, Randeva HS.
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