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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

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Images

SDS-PAGE - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177), expandable thumbnail
  • Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177), expandable thumbnail
  • Functional Studies - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177), expandable thumbnail
  • Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177), expandable thumbnail
  • Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (AB219177), expandable thumbnail

Publications

Key facts

Assay time
45m
Sample type
Adherent cells, Suspension cells, Tissue

What's included?

100 Test
Components
200X DTT
1 x 1 mL
Cytoplasm Extraction Buffer
1 x 55 mL
Nuclear Extraction Buffer
1 x 55 mL
Nuclear Lysis Buffer
1 x 55 mL
Protease Inhibitor Cocktail
1 x 1 mL

Recommended products

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

Key facts

Assay time
45m
Sample type
Adherent cells, Suspension cells, Tissue

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
Multi
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols

Notes

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.

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • SDS-PAGE - Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177), expandable thumbnail

    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.

  • Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177), expandable thumbnail

    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.

  • Functional Studies - Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177), expandable thumbnail

    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) (Histone Deacetylase (HDAC) Activity Assay Kit (Fluorometric) ab156064). Extract from of 3 x 104 cells was used per assay test.

  • Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177), expandable thumbnail

    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.

  • Western blot - Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177), expandable thumbnail

    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.

  • Sandwich ELISA - Nuclear Extraction Kit - Soluble / Insoluble Fractions (ab219177), expandable thumbnail

    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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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