Rat Brain Whole Tissue Lysate (Post-Natal Whole)

Rat

ELISA, Immunoprecipitation, SDS-PAGE, Western Blot

This product has been tested by SDS-PAGE. Multi-purpose Rat brain, post natal, Whole Cell Lysate is especially prepared as positive control for multiple assays including western blot, immunoprecipitation (IP), capture ELISA or other assays requiring native protein sample. For separation by SDS-PAGE and subsequent western blot analysis, lysates should be diluted by user to desired concentration in SDS-PAGE buffer with 2-mercaptoethanol or dithiothreitol as the reducing agent and heated to 95C for 5 minutes. Sample is ready for use in immunoprecipitation and ELISA experiments, conditions should be optimized by the user. It is recommended to use TrueBlot IP reagents for immunoprecipitation experiments.

Tissue

Post-Natal

Whole

Tissues were washed exhaustively with PBS to remove blood and other debris. A lysate was prepared by homogenizing the tissue and washing the cells in cold PBS. Washed cells were incubated at 4C in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, and 0.5% sodium deoxycholic acid the cells. Protein integrity is ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 uM Aprotinin, 5 uM Bestatin, 1.5 uM E-64, 2 uM Leupeptin Hemisulfate and 1 uM Pepstatin A). The following phosphatase inhibitors were also added: 1 mM NaF and 1 mM Na3VO4. Cell debris was removed by centrifugation and membrane filtration. Protein concentration was determined by Lowry assay using a commercially available kit. The protein concentration was adjusted to 1 mg/ml using RIPA buffer.

Multi-step

1X RIPA Buffer with HALT Protease and Phosphatase Inhibitors

No Preservative

Please contact technical services for concentration.

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Store at -70C. Avoid freeze-thaw cycles.