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. 2015 Sep 28;19(12):2865–2873. doi: 10.1111/jcmm.12679

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© 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Subcellular Distribution of OLFM4 in Neutrophils. Neutrophils (4.8 × 108) isolated from peripheral blood were cavitated in 12 ml disruption buffer and 5 ml of the post‐nuclear supernatant applied to each of two 4‐layer Percoll density gradients. Separation of organelles was obtained by high‐speed centrifugation and 1 ml fractions were collected from the bottom of the centrifuge tube, pooled from the two identical gradients and assayed for myeloperoxidase (MPO; azurophil granules), NGAL (specific granules), Gelatinase (gelatinase granules), Albumin (secretory vesicles) and human leucocyte antigen (HLA; plasma membranes) and OLFM4. The distribution profile of OLFM4 is identical to the profile of the specific granule protein NGAL.