CelLytic™ MT Cell Lysis Reagent

Protocol tips

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	- The prostate tissues of rats are homogenized using cold CelLytic™ MT Mammalian Tissue Lysis/Extraction Reagent (sigma) containing 2 mM sodium orthovanadate, 1 mM PMSF and protein cocktail inhibitor (1X, Sigma). The homogenate is centrifuged at 10,000 rpm for 10 min.

Protocol tips
- The prostate tissues of rats are homogenized using cold CelLytic™ MT Mammalian Tissue Lysis/Extraction Reagent (sigma) containing 2 mM sodium orthovanadate, 1 mM PMSF and protein cocktail inhibitor (1X, Sigma). The homogenate is centrifuged at 10,000 rpm for 10 min.

Publication protocol

The prostate and bladder tissues of rats injected with formalin or saline (n = 6 per group) were homogenized using cold CelLytic™ MT Mammalian Tissue Lysis/Extraction Reagent (sigma) containing 2 mM sodium orthovanadate, 1 mM PMSF and protein cocktail inhibitor (1X, Sigma). The homogenate was centrifuged at 10,000 rpm for 10 min and the resulting supernatants were stored at -80 °C until assayed. 28 proteins including interleukins IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-13, IL-17A and IL-18; CXC chemokines (CXCL1, CXCL2, CXCL5 and CXCL10), CC chemokines (CCL2, CCL3, CCL5); Growth factors NGF, BDNF, VEGF and G-CSF, other inflammatory mediators such as eotaxin, leptin, IFN-γ and TNFα levels were determined on a Luminex 100 using a MILLIPLEX MAP Rat Cytokine/Chemokine Panel (Millipore, Billerica, MA). Levels of NGF and BDNF were determined using individual ELISA kits procured from Promega, USA. Protein estimation was done by BCA Protein Assay Kit (Pierce, Rockford, Illinois) to standardize the chemokine concentrations relative to tissue protein levels, which are expressed as pg/mg of total protein.

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Manufacturer protocol

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