Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177)
Key features and details
- Assay time: 45 min
- Sample type: Adherent cells, Suspension cells, Tissue
Overview
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Product name
Nuclear Extraction Kit - Soluble / Insoluble Fractions
See all Nuclear Extraction kits -
Sample type
Tissue, Adherent cells, Suspension cells -
Assay time
0h 45m -
Product overview
Nuclear extraction kit ab219177 offers a simple 45 minute method for extracting nuclear proteins from mammalian cells or tissues.
For optimal extraction of nuclear proteins, the kit produces three distinct extracts (fractions): soluble nuclear proteins (nuclear extract 1), nuclear proteins that remain insoluble after production of nuclear extract 1 (nuclear extract 2), and cytoplasmic proteins.
The kit contains reagents for 100 extractions, where each extraction starts with 5 x 106 cells or 50 mg of tissue. Typical yields from each extraction are 0.3 – 0.4 mg of soluble nuclear proteins, 0.1 – 0.2 mg of insoluble nuclear proteins and 0.6 – 0.7 mg of cytoplasmic proteins. Nuclear proteins extracted with this kit can be used in a variety of applications such as western blotting and nuclear enzyme assays.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 100 tests 200X DTT 1 x 1ml Cytoplasm Extraction Buffer 1 x 55ml Nuclear Extraction Buffer 1 1 x 55ml Nuclear Extraction Buffer 2 1 x 55ml Protease Inhibitor Cocktail 1 x 1ml
Images
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SDS PAGE analysis of HeLa cell fractionation using Nuclear Extract Kit ab219177.
Extracts were prepared from 5 x 106 HeLa (human epithelial cell line from cervix adenocarcinoma) cells using Nuclear Extract Kit ab219177 and analysed by SDS-PAGE with Coomasie Blue staining. Extract or whole cell lysate from 3 x 104 cells was loaded in each lane along with BSA standards. Lanes: 1. MW marker; 2. Whole cell lysate; 3. Cytoplasmic extract; 4. Nuclear extract 1; 5. Nuclear extract 2; 6. MW marker; 7. BSA, 2.0 µg; 8. BSA 1.0 µg; 9. BSA 0.5 µg; 10. BSA 0.2 µg; 11. BSA 0.1 µg.
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Comparison of HeLa cell fractionation by Nuclear Extract Kit ab219177 and leading commercially available nuclear fractionation kits.Extracts were prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells using the manufacturer’s protocol and analyzed by Western blot. For each manufacturer: (1) cytoplasmic extract, (2) nuclear extract, (3) nuclear extract 2 or resolubilized insoluble nuclear fraction.
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Measurement of Histone Deacetylase (HDAC) activity in HeLa cell fractions prepared using Nuclear Extract Kit ab219177.Extracts were prepared from 5 x 106 HeLa (human epithelial cell line from cervix adenocarcinoma) cells using Nuclear Extract Kit ab219177. Note that protease inhibitors were omitted from all buffers to prevent interference with the HDAC activity assay. The enzymatic activity of HDACs in each fraction was measured using the Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) (ab156064). Extract from of 3 x 104 cells was used per assay test.
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Comparison of nuclear extraction from different cell lines.Extracts were prepared from 5 x 106 cells from RAJI (human lymphoblastoid cell line), HEK-293 (human embryonic kidney cell line), SHSY-5Y (human neuroblastoma cell line) and U87-MG (human glioblastoma cell line) cell lines using Nuclear Extract Kit ab219177 and analysed by Western blot. Extract from 3 x 104 cells was loaded per lane.
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Comparison of nuclear extraction from different tissues.Extracts were prepared from 120 mg of mouse tissue using Nuclear Extract Kit ab219177 and analysed by Western blot. Extract from 0.4 mg of tissue was loaded per lane.