Nuclear extraction kit ab219177 provides a simple method for extracting nuclear proteins from mammalian cells or tissues in 45 minutes. The extracted protein fractions are suitable for western blot and functional assays.
- Used to extract soluble nuclear proteins, insoluble nuclear proteins and cytoplasmic proteins from adherent cells, suspension cells and tissue samples
- Kit contains reagents for 100 extractions
Nuclear extraction kit ab219177 provides a simple method for extracting nuclear proteins from mammalian cells or tissues in 45 minutes. The extracted protein fractions are suitable for western blot and functional assays.
- Used to extract soluble nuclear proteins, insoluble nuclear proteins and cytoplasmic proteins from adherent cells, suspension cells and tissue samples
- Kit contains reagents for 100 extractions
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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Terms & Conditions.
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.
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.
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) (Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) ab156064). Extract from of 3 x 104 cells was used per assay test.
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.
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.
NFkB p50 activity in treated and untreated Hela nuclear extract.
Nuclear extracts from Hela cells and Hela cells treated with 25 ng/mL TNF alpha were assayed from 0.1875 µg-12 µg/well using the Abcam NFkB p50 Transcription Factor Activity Assay. Data shown is from wells assayed in duplicate. Comparison of treated to untreated Hela nuclear extract shows average fold change ~17. ab219177 Nuclear Extraction kit was used to extract nuclear proteins.
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