When talking about a chemokine receptor CXCR4, I expect membrane/cytoplasmatic staining, but when using different pretreatments, I find a different staining pattern. I used heat retrieval and got nuclear staining, and when using Protease pretreatment I get cytoplasmatic staining in normal tissues. Could it be the structure of the receptor changing in different pretreatments, or has anyone else found the same problems? In literature, I found different articles using the same clone antibody but also different results, one paper used protease pretreatment and found cytoplasmatic staining, and the other used heat retrieval and found both staining patterns or only nuclear.
1. Kato, M., Kitayama, J., Kazama, S., and Nagawa, H. 2003. Expression pattern of CXC chemokine receptor-4 is correlated with lymph node metastasis in human invasive ductal carcinoma. Breast Cancer Res 5:R144-150.
2. Woo, S.U., Bae, J.W., Kim, C.H., Lee, J.B., and Koo, B.W. 2008. A significant correlation between nuclear CXCR4 expression and axillary lymph node metastasis in hormonal receptor negative breast cancer. Ann Surg Oncol 15:281-285.
3. Kang, H., Watkins, G., Parr, C., Douglas-Jones, A., Mansel, R.E., and Jiang, W.G. 2005. Stromal cell derived factor-1: its influence on invasiveness and migration of breast cancer cells in vitro, and its association with prognosis and survival in human breast cancer. Breast Cancer Res 7:R402-410.
Don-Salu-Hewage AS, Chan SY, McAndrews KM, Chetram MA, Dawson MR, et al. (2013) Cysteine (C)-X-C Receptor 4 Undergoes Transportin 1- Dependent Nuclear Localization and Remains Functional at the Nucleus of Metastatic Prostate Cancer Cells. PLoS ONE 8(2): e57194. doi:10.1371/ journal.pone.0057194
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