||Detergents typically present in cell lysis buffers are thought to disrupt organelles and compartments and increase the exposure of soluble phospho-proteins to phosphatases and proteases thereby resulting in uncontrolled dephosphorylation and proteolysis.
Phospho-Sure™ RTD Neuronal extraction buffer is optimized for the extraction of phosphorylated proteins from neuronal and other soft tissues types. The buffer extracts the phosphoproteins in a native state without the use of harsh detergents or oxidizers, and it is specially formulated to help maintain phosphoproteins and protect them from degradation better than traditional detergent based extraction buffers.
||See product label
||Please download the protocol below for detailed instructions on how to use Phospho-Sure in neuronal and other soft tissues.
||Refer to the publication by Suneja et al (2006) where this buffer was shown to improve recovery of phospho-proteins such as ERK1/2-P and SAP/JNK-P in the dissection and homogenization of guinea pig brain tissue.
||Phospho-Sure™ RTD™ is designed as both an isolation and extraction buffer and for best results the intact tissue should be bathed in ice-cold Phospho-sure™ RTD™ buffer during excision of the tissue. Once the desired tissue piece is isolated in Phospho-sure™ RTD™ it may be transferred to a small volume of ice-cold Phospho-Sure™ RTD™ for disruption.
||Powder containing no preservatives. Preparation of Phospho-Sure™ RTD™ is very simple. Simply add sterile ddH2O directly to the powdered solution in the supplied bottle, swirl briefly to dissolve the chemicals and use. The material contains no preservatives so sterile technique should always be used.
||Add 186 ml of sterile double distilled water and mix by gently swirling, final volume 200 ml at 20°C.
||The dry, unopened container should be stored at room temperature in a dry or desiccated location protected from light. Do not store in the refrigerator unless material is in a dry, moisture free environment. Material is hydroscopic so once the seal is broken it should be hydrated and not resealed while dry. Once hydrated, the buffer can be stored at 4°C for up to 3 months. Solution can be frozen but clumping may occur upon thawing and is not recommended.
||Powdered format can be stored up to 12 months after purchase under cool, dry conditions.
||1. Suneja SK, Mo Z, Potashner SJ. (2006) Phospho-CREB and other phospho-proteins: improved recovery from brain tissue.
J Neurosci Methods. 2006 Jan 30;150(2):238-41.
2. Elvira Mass, Dagmar Wachten, Anna C. Aschenbrenner, André Voelzmann, Michael Hochemail (2014) Murine Creld1 Controls Cardiac Development through Activation of Calcineurin/NFATc1 Signaling, Developmental Cell Volume 28, Issue 6, p711–726