Nuclear Extraction Kit - Soluble / Insoluble Fractions

• SDS-PAGE

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SDS-PAGE - Nuclear Extraction Kit - Soluble / Insoluble Fractions (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.

• FuncS

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Functional Studies - Nuclear Extraction Kit - Soluble / Insoluble Fractions (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.

• sELISA

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Sandwich ELISA - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177)

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.

• WB

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Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177)

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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• WB

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Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177)

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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• WB

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Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177)

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.

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