使用说明：
对于培养细胞样品：
1. 融解RIPA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150-250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150-250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。
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和免疫沉淀等操
   作。
   裂解液用量说明：通常6孔板每孔细胞加入150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200微升或250微升。
对于组织样品：
1. 把组织剪切成细小的碎片。
2. 融解RIPA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150-250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和免疫沉淀等操
   作。
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。
注：RIPA裂解液的裂解产物中经常会出现一小团透明胶状物，属正常现象。该透明胶状物为含有基因组
    DNA等的复合物。在不检测和基因组DNA结合特别紧密的蛋白的情况下，可以直接离心取上清用于后
    续实验；如果需要检测和基因组结合特别紧密的蛋白，则可以通过超声处理打碎打散该透明胶状
    物，随后离心取上清用于后续实验。如果检测一些常见的转录因子，例如NF-kappaB、p53等时，通
    常不必进行超声处理，就可以检测到这些转录因子。

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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75.Hao M, Kuang HY, Fu Z, Gao XY, Liu Y, Deng W
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76.Chen J, Lin M, Li N, Lin L, She F
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77.Wang SB, Shi Q, Xu Y, Xie WL, Zhang J, Tian C, Guo Y, Wang K, Zhang BY, Chen C, Gao C,
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80.Zhang Y, Zhou L, Zhang X, Bai J, Shi M, Zhao G.
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82.Yang XH, Wang YH, Wang JJ, Liu YC, Deng W, Qin C, Gao JL, Zhang LY.
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83.Jia LQ, Yang GL, Ren L, Chen WN, Feng JY, Cao Y, Zhang L, Li XT, Lei P.
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84.Wang J, Sun LL, Zhu WL, Sun Y, Liu JF, Lu L, Shi J
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85.Wang D, Tang F, Wang S, Jiang Z, Zhang L
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86.Yu L, Liu Y, Qiu Z, Liu S, Gao X, Zhu D.
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87.Li J, Guo Y, Liang X, Sun M, Wang G, De W, Wu W
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88.Liu K, Wang H, Long Y, Ye J, Yuan L
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91.Yu X, Tang J, Wang Q, Ye W, Tao K, Duan S, Lu C, Yang X, Dong S, Zheng X, Wang Y
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92.Wang Y, Liu Q, Liu Z, Li B, Sun Z, Zhou H, Zhang X, Gong Y, Shao C
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93.Liu Y, Xiao Z, Yang D, Ren L, Liu G, Yang L
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94.Ding D, Xu H, Liang Q, Xu L, Zhao Y, Wang Y
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95.Li R, Yu C, Gao R, Liu X, Lu J, Zhao L, Chen X, Ding Y, Wang Y, He J.
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97.Wang C, Zhang L, He Q, Feng X, Zhu J, Xu Z, Wang X, Chen F, Li X, Dong J.
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98.Xue B, Wu W, Huang K, Xie T, Xu X, Zhang H, Qi C, Ge J, Yu Y.
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100.Li G, Zhang S, Fang H, Yan B, Zhao Y, Feng L, Ma X, Ye X.
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101.Zhang X, Chen G, Lu Y, Liu J, Fang M, Luo J, Cao Q, Wang X.
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102.Chen X, Chen L, Zhang R, Yi Y, Ma Y, Yan K, Jiang X, Wang X.
   ADAM17 regulates self-renewal differentiation of U87 glioblastoma stem cells.
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103.Ye HH, Wu KJ, Fei SJ, Zhang XW, Liu HX, Zhang JL, Zhang YM.
   Propofol participates in gastric mucosal protection through inhibiting the toll-like
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104.Zheng Y, He L, Wan Y, Song J.
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105.Liu X, Wang M, Chen H, Guo Y, Ma F, Shi F, Bi Y, Li Y.
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106.Liang X, Cheng S, Jiang X, He X, Wang Y, Jiang Z, Hou W, Li S, Liu Y, Wang Z.
   The noncircadian function of the circadian clock gene in the regulation of male
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107.Wei Q, Ren X, Jiang Y, Jin H, Liu N, Li J.
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108.Shou Y, Yang Y, Xu MS, Zhao YQ, Ge LB, Zhang BM.
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109.Gao L, Song JR, Zhang JW, Zhao X, Zhao QD, Sun K, Deng WJ, Li R, Lv G, Cheng HY,
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   Chloroquine promotes the anticancer effect of TACE in a rabbit VX2 liver tumor model.
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110.Zheng XF, Li B, Zhang YH, Yang YH, Meng XY, Jiang SD, Jiang LS.
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111.Liu W, Otkur W, Li L, Wang Q, He H, Zang L, Hayashi T, Tashiro S, Onodera S, Xia M,
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112.Yuan L, Zhang L, Dong X, Zhao H, Li S, Han D, Liu X.
   Apoptin selectively induces the apoptosis of tumor cells by suppressing the
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113.Pang X, Wei Y, Zhang Y, Zhang M, Lu Y, Shen P.
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114.Xu J, Jia YY, Chen SR, Ye JT, Bu XZ, Hu Y, Ma YZ, Guo JL, Liu PQ.
   (E)-1-(4-ethoxyphenyl)-3-(4-nitrophenyl)-prop-2-en-1-one suppresses LPS-induced
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115.Xu P, Zhang XX, Miao C, Fu Z, Li Z, Zhang G, Zheng M, Liu Y, Yang L, Wang T.
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116.Han J, Ding R, Zhao D, Zhang Z, Ma X.
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117.Jin J, Zhao WC, Yuan F.
   CXCR7/CXCR4/CXCL12 Axis Regulates the Proliferation, Migration, Survival Tube
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118.Wang Q, Li LH, Gao GD, Wang G, Qu L, Li JG, Wang CM.
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119.Wang J, Wang X, Wu G, Hou D, Hu Q.
   MiR-365b-3p, down-regulated in retinoblastoma, regulates cell cycle progression
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110.Wang J, Huang H, Wang C, Liu X, Hu F, Liu M.
   MicroRNA-375 sensitizes tumour necrosis factor-alpha (TNF-α)-induced apoptosis in head
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121.Li A, Lu D, Zhang Y, Li J, Fang Y, Li F, Sun J.
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122.Mu H, Yao RB, Zhao LJ, Shen SY, Zhao ZM, Cai H.
   Sinomenine Decreases MyD88 Expression Improves Inflammation-Induced Joint Damage
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123.Ru R, Yao Y, Yu S, Yin B, Xu W, Zhao S, Qin L, Chen X.
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124.Xu D, Zhang T, Chen X, Zhou Q, Liu C, Deng Z, Zhang L, Ying C, Zhang W, Gu M.
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125.Zhang RX, Yan XB, Gu YH, Huang D, Gan L, Han R, Huang LH.
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126.Li H, Zhou RJ, Zhang GQ, Xu JP.
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127.Mo QQ, Chen PB, Jin X, Chen Q, Tang L, Wang BB, Li KZ, Wu P, Fang Y, Wang SX, Zhou JF,
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   Inhibition of Hec1 expression enhances the sensitivity of human ovarian cancer cells to
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128.Li L, Han M, Li S, Wang L, Xu Y.
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129.Zhu X, Jiang Y, Shan PF, Shen J, Liang QH, Cui RR, Liu Y, Liu GY, Wu SS, Lu Q, Xie H,
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130.Xu N, Yuan H, Liu W, Li S, Liu Y, Wan J, Li X, Zhang R, Chang Y.
   Activation of RAW264.7 mouse macrophage cells in vitro through treatment with
   recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB
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131.Liu L, Liu L, Bo T, Li S, Zhu Z, Cui R , Mao D.
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132.Zhang J, Ni SS, Zhao WL, Dong XC, Wang JL.
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133.Fei X, Jiang S, Zhang S, Li Y, Ge J, He B, Goldstein S, Ruiz G.
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134.Ding F, Yu L, Wang M, Xu S, Xia Q, Fu G.
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135.Sun LL, Zhang Y, Liu JF, Wang J, Zhu WL, Zhao LY, Xue YX, Lu L, Shi J.
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136.Peng H, Zhang W, Xiao T, Zhang Y.
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137.Guan X, Guan Y.
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138.Cong ZX, Wang HD, Wang JW, Zhou Y, Pan H, Zhang DD, Zhu L.
   ERK PI3K signaling cascades induce Nrf2 activation regulate cell viability
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139.Shen R, Xie F, Shen H, liu Q, Zheng T, Kou X, Wang D, Yang J.
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140.Liu E, Kuang Y, He W, Xing X, Gu J.
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141.Ge D, Kong X, Liu W, Zhao J, Su L, Zhang S, Zhang Y, Zhao B, Miao J.
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142.Yang J, Huang H, Yang H, He X, Jiang X, Shi Y, Alatangaole D, Shi L, Zhou N.
   Specific activation of the G protein-coupled receptor BNGR-A21 by the neuropeptide
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144.Zhao Y, Wang MY, Hao K, Chen XQ, Du JZ.
   CRHR1 mediates p53 transcription induced by high altitude hypoxia through ERK 1/2
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146.Sun LF, Shi C, Yuan L, Sun Y, Yao XX, Ma JW, Huang CM, Zhu HF, Lei P, Shen GX.
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147.Chen Z, Yang Y, Yang X, Zhou C, Li F, Lei P, Zhong L, Jin X, Peng G.
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148.Fu Y, Zhang P, Ge J, Cheng J, Dong W, Yuan H, Du Y, Yang M, Sun R, Jiang H.
   Histone deacetylase 8 suppresses osteogenic differentiation of bone marrow stromal cells
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149.Wang C, Pan Y, Zhang R, Bai B, Chen J, Randeva HS.
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150.Sun H, Wu Y, Fu D, Liu Y, Huang C.
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151.Xie Q, Wang Z, Bi X, Zhou H, Wang Y, Gu P, Fan X.
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154.Jiang T, Yu JT, Zhu XC, Tan MS, Gu LZ, Zhang YD, Tan L.
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155.Wang G, Liu X, Meng L, Liu S, Wang L, Li J, Cui C, Meng J, Hu S, Wei Y.
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156.Tang W, Yang J, Zhang F, Guo H, Peng F, Wang X.
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157.Deng Z, Yan F, Jin Q, Li F, Wu J, Liu X, Zheng H.
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158.Zhu XC, Jiang T, Zhang QQ, Cao L, Tan MS, Wang HF, Ding ZZ, Tan L, Yu JT.
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159.Guo F, Jiang T, Song W, Wei H, Wang F, Liu L, Ma L, Yin H, Wang Q, Xiong L.
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160.Wu H, Wang K, Liu W, Hao Q.
   PTEN overexpression improves cisplatin-resistance of human ovarian cancer cells through
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161.Guo S, Xu X, Tang Y, Zhang C, Li J, Ouyang Y, Ju J, Bie P, Wang H.
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162.Xu J, Ruan L, Shi H.
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163.Cong ZX, Wang HD, Zhou Y, Wang JW, Pan H, Zhang DD, Zhang L, Zhu L.
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