FAQs for Liver Lysate (NB820-59232). (Showing 1 - 2 of 2 FAQs).
Which fraction is contained in this lysate? Also, on the datasheet, the applications of this lysate include enzyme activity analysis - what does this mean? Lastly, could you tell me if using RIPA buffer affects the enzyme activity or not?
The adult normal human liver whole tissue lysate with catalog number NB820-59232 is prepared from whole tissue homogenates and is stored in HEPES, pH 7.9, MgCl2, KCl, EDTA, Sucrose, Glycerol, Detergent with a cocktail of protease inhibitors. As such, the activity of the expressed enzymes should be preserved. During the Quality Control process the protein lysate is verified on SDS-PAGE analysis as a consistent pattern comparing to a standard protein lysate. This uses the following biomarkers: GAPDH for total protein, cytoplasmic and universal total protein; Histone for nuclear protein; Na/K ATPase for membrane protein; Vimentin for cytoskeleton protein. NB820-59232 is validated for Western blotting and immunoprecipitation, and is guaranteed for 6 months from the date of receipt. We recommend that you store this product at -80C and avoid freeze-thaw cycles.
Regarding Human Liver Whole Tissue Lysate (Adult Whole Normal) NB820-59232 and Human Liver Whole Tissue Lysate (Adult Whole Liver Cirrhosis):1. Does the lysate also contain the membrane protein? If yes, will it be possible to isolate the membrane fraction from the lysate? For my experiment I segregate the cytosolic and membrane fraction.2. Do you treat , Human Liver Whole Tissue Lysate (Adult Whole Liver Cirrhosis) NB820-59328, with DNase1 to avoid coagulation while heating prior to loading the lysate in SDS running gel? If not DNase1, what you use to avoid the coagulation?
All Whole Tissue Lysates are prepared using the following kit:https://www.novusbio.com/products/total-protein-kit_nbp2-37853We don’t treat the lysates with DNase. We do not notice any coagulation when loading on an SDS-PAGE gel. Just heating the sample in standard loading buffer works well. The whole tissue lysates should contain plasma membrane fragments, however some components (e.g. mitochondria, lysosomes) will probably precipitate in the final 18,000g spin.We do however have corresponding membrane fractions:Normal: https://www.novusbio.com/products/liver-lysate_nb820-60598Normal, left lobe: https://www.novusbio.com/products/liver-lysate_nb820-60599Cirrhosis: https://www.novusbio.com/products/liver-lysate_nbl1-26449
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