101110 26 1 Select tissue livertripe 2 Pulverise tissue to get crude soluble

101110 26 1 select tissue livertripe 2 pulverise

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10/11/10 26 1. Select tissue: liver/tripe 2. Pulverise tissue to get crude soluble protein extract; centrifuge 3. Parallel dilution; make standard purified (SPP) dilution series (Standard curve) 4. Measure absorbance of diluted extract; calculate concentration of extract using standard curve Steps in Protein Extraction
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Electrophoresis and Electroblotting of Proteins 14 10/11/10 27 1. Select tissue: liver/tripe 2. Pulverise tissue to get crude soluble protein extract; centrifuge 3. Parallel dilution; make standard purified (SPP) dilution series (Standard curve) 4. Measure absorbance of diluted extract; calculate concentration of extract using standard curve Steps in Protein Extraction 10/11/10 28 Smooth ER Glycogen granules Rough Endoplasmic Reticulum Liver Tissue Characteristics
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Electrophoresis and Electroblotting of Proteins 15 10/11/10 29 Liver functions Storage of glycogen, ions, lipids, etc. Processing, detoxification and storage of molecules from intestines Synthesis of blood plasma proteins Generation of body heat Breakdown of RBC & Bile formation 10/11/10 30 Liver & Onions Recipe “In a small mixing bowl combine together chopped marjoram, thyme, parsley flakes, seasoned salt, prepared mustard and black pepper. Crush and mix all of the ingredients together to form your seasoning mixture. After about 40 minutes, remove sliced liver from milk and pat dry with paper towels. Coat each piece of liver first with seasoning mixture and then with flour. Heat vegetable oil in a large heavy skillet over medium-high heat. Fry the liver to your desired doneness. Remove cooked liver and place on a warmed plate until onions are done. Place onions into skillet, season with salt and pepper and brown, turning occasionally. Serve with steamed rice, vegetables and cornbread.” http:// liver-and-onions-recipe.html
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Electrophoresis and Electroblotting of Proteins 16 10/11/10 31 The Omasum, “tripe” (third division of a ruminant’s stomach) Exterior Interior 10/11/10 32 Tripe Secretory layer Muscle layer Arteries &Veins Connective tissue d0.biggestmenu.com/.../ 20/b93d008311646a0f_m.jpg
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Electrophoresis and Electroblotting of Proteins 17 10/11/10 33 1. Select tissue: liver/tripe 2. Pulverise tissue to get crude soluble protein extract; centrifuge 3. Parallel dilution; make standard purified (SPP) dilution series (Standard curve) 4. Measure absorbance of diluted extract; calculate concentration of extract using standard curve Steps in Protein Extraction 10/11/10 34 centrifuge Extract total proteins from tissues or organs to prepare crude extracts crushed cells Pellet = intact cells, cell debris Supernatant = “ crude extract ”, i.e., soluble proteins, plus DNA and RNA and other soluble molecules, plus membrane vesicles containing membrane proteins.
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