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ER0100

Sigma-Aldrich

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

Quality Level

technique(s)

fractionation: suitable

storage temp.

2-8°C

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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Hazard Classifications

Eye Irrit. 2

Storage Class Code

10 - Combustible liquids

WGK

WGK 1


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Yuan Tian et al.
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The rate of behavioural decline in the ageing population is remarkably variable among individuals. Despite the considerable interest in studying natural variation in ageing rate to identify factors that control healthy ageing, no such factor has yet been found. Here
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Resistance to BRAF and MEK inhibitors (BRAFi + MEKi) in BRAF-mutant tumors occurs through heterogeneous mechanisms, including ERK reactivation and autophagy. Little is known about the mechanisms by which ERK reactivation or autophagy is induced by BRAFi + MEKi. Here
Liou Y Sun et al.
eLife, 2, e01098-e01098 (2013-11-01)
We examine the impact of targeted disruption of growth hormone-releasing hormone (GHRH) in mice on longevity and the putative mechanisms of delayed aging. GHRH knockout mice are remarkably long-lived, exhibiting major shifts in the expression of genes related to xenobiotic
Leilei Zhang et al.
Cancer research, 79(16), 4072-4085 (2019-06-23)
ME-344 is a second-generation isoflavone with unusual cytotoxic properties that is in clinical testing in cancer. To identify targets that contribute to its anticancer activity and therapeutic index, we used lung cancer cell lines that are naturally sensitive or resistant

Articles

The isolation of subcellular fractions by centrifugation is a commonly used technique and is widely applicable across multiple cell and tissue types. Because organelles differ in their size, shape, and density, centrifugation can be easily employed to separate and purify organelle fractions from gently homogenized samples.

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