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On past January 21 we received from you a dosis of the anti human HLA E monoclonal antibody (MEM-E/06) that we purchased aiming to monitor the surface expression of HLA E in cells infected with different viruses and uninfected cell controls. We performed flow cytometry labeling B lymphoblastoid and epithelial human cells with MEM-E in parallel with moab anti human HLA I A, B, C W6/32 from Serotec. The labelling pattern in the uninfected cells were similar for MEM-E/06 and W&/32, ( we can show you the Fascan profiles) and we were unable to monitor changes in HLA E expression upon virus infections. As you state in the MEM-E/06 datasheet it has been recently reported that this moab crossreact broadly with several HLA A, B and C. Could you please send me any suggestion to overcome this trouble? Thanking in advance your cooperation,
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ANSWER: |
Thank you for your enquiry. We have sold several vials of this product and do not have any complaint. This is a very specific question. We would suggest, (together with FACS analysis) to lyse the same cells in native lysis buffer (1%NP-40 plus inhibitors) and then subjected to non-denaturing, non-reducing SDS-PAGE (even boiling should be omitted). Try to probe the WB blots with Ab2216 MEM-E/02 (absolutely HLA-E specific) and compare with W6/32 to see changes? We are going to release a new anti HLA-E antibodies for flow cytometry which is much more specific (very restricted cross-reactivity).
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On past January 21 we received from you a dosis of the anti human HLA E monoclonal antibody (MEM-E/06) that we purchased aiming to monitor the surface expression of HLA E in cells infected with different viruses and uninfected cell controls. We performed flow cytometry labeling B lymphoblastoid and epithelial human cells with MEM-E in parallel with moab anti human HLA I A, B, C W6/32 from Serotec. The labelling pattern in the uninfected cells were similar for MEM-E/06 and W&/32, ( we can show you the Fascan profiles) and we were unable to monitor changes in HLA E expression upon virus infections. As you state in the MEM-E/06 datasheet it has been recently reported that this moab crossreact broadly with several HLA A, B and C. Could you please send me any suggestion to overcome this trouble? Thanking in advance your cooperation,
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ANSWER: |
Thank you for your enquiry. We have sold several vials of this product and do not have any complaint. This is a very specific question. We would suggest, (together with FACS analysis) to lyse the same cells in native lysis buffer (1%NP-40 plus inhibitors) and then subjected to non-denaturing, non-reducing SDS-PAGE (even boiling should be omitted). Try to probe the WB blots with Ab2216 MEM-E/02 (absolutely HLA-E specific) and compare with W6/32 to see changes? We are going to release a new anti HLA-E antibodies for flow cytometry which is much more specific (very restricted cross-reactivity).
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