The Endoplasmic Reticulum Isolation Kit does not contain membrane proteins and peripheral membrane proteins. The kit includes the following components:
• Isotonic Extraction Buffer 5× (100 mL): Contains 50 mM HEPES (pH 7.8), 1.25 M sucrose, 5 mM EGTA, and 125 mM KCl.
• Hypotonic Extraction Buffer 10× (10 mL): Contains 100 mM HEPES (pH 7.8), 10 mM EGTA, and 250 mM KCl.
• Calcium Chloride Solution (5 mL): A 2.5 M CaCl2 solution.
• OptiPrep™ Density Gradient Medium (100 mL): A 60% (w/v) solution of iodixanol in water.
• Needle (4 inch, 20 gauge): One each.
ER0100
Endoplasmic Reticulum Isolation Kit
isolation of intact ER from mammalian soft tissues and cultured cells
Synonym(s):
ER isolation kit
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About This Item
UNSPSC Code:
12352200
NACRES:
NA.32
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General description
The endoplasmic reticulum (ER) is a network of interconnected tubules, vesicles, and sacs. It plays a role in specialized cellular functions including protein synthesis, sequestering of calcium, production of steroids, storage and production of glycogen, and insertion of membrane proteins.
Application
The Endoplasmic Reticulum Isolation Kit contains all the reagents required for ER preparations with various degrees of purity: crude ER (microsomes), Ca2+ precipitated rough ER (RER) enriched microsomes, and density gradient purified rough ER (RER) and smooth ER (SER). The ER isolation process can be monitored by measuring the activity of NADPH cytochrome c reductase using the Cytochrome c Reductase Assay Kit, (Cat. No. CY0100). The enzyme is an ER membrane protein that is commonly used as an ER marker. The isolated ER can be used for studying the cytochrome P450 system and xenobiotic metabolism, for studying lipid metabolism, and for the recovery of ER membrane and lumenal proteins. The kit was tested with rat liver, kidney, and brain; mouse liver; rabbit liver, kidney, spleen, heart, and brain; and with Jurkat and HeLa cell lines.
The Endoplasmic Reticulum Isolation Kit is used for the efficient enrichment of functional endoplasmic reticulum (ER) from mammalian tissue or cell culture. The isolated ER are useful for protein synthesis, degradation and metabolic pathway analysis. Metabolic abnormalities of ER include carnitine acetyltransferase deficiency in children, cystic fibrosis and many more ER protein retention disorders. Suitable for up to 50 g of tissue or ~20 mL of packed tissue.
Features and Benefits
- Specially formulated extraction reagents for research scale applications - save time and minimize waste
- Includes a calcium chloride solution - for quick and simple precipitation of rough ER without need for ultracentrifugation
- Produces functional, intact organelles - resulting ER are suitable for functional studies, lipid metabolism, and protein profiling
- Compatible with products for structure confirmation - easily confirm intactness with companion kit test for cytochrome C reductase (CY0100)
- Can be used to isolate ER from soft animal tissues and cultured cells
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Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
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Articles
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
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Does the Endoplasmic Reticulum (ER) obtained with the Endoplasmic Reticulum Isolation kit contain membrane proteins and peripheral membrane proteins?
1 answer-
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What level of purity of endoplasmic reticulum (ER) can be anticipated when utilizing the Endoplasmic Reticulum Isolation kit with low, medium, or ultra centrifugation?
1 answer-
When using low and medium-speed centrifugation steps, the resulting preparations are generally expected to be crude. To achieve an expectation of purity, it is advisable to follow the ultracentrifugal procedure. Notably, during quality control, no specific purity specifications were found. The Specification Sheet merely indicates that the kit must be suitable. Purity levels may fluctuate based on factors such as tissue type, specimen age, and other variables. If there are concerns about mitochondrial or other impurities, it would be prudent to test for mitochondrial markers in the collected ER fraction. Additionally, the level of purity is likely to vary depending on the carefulness with which the different fractions are collected.
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