ab221031 Histone Extraction Kit - Rapid / Ultra-pure enables the extraction of core histones from mammalian cells or tissues in 1 hour using a simple procedure with 20 minutes hands-on time.
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- Molecular neurobiology 56:4549-45652018Resveratrol Preconditioning Induces Genomic and Metabolic Adaptations within the Long-Term Window of Cerebral Ischemic Tolerance Leading to Bioenergetic Efficiency.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNathalie Khoury,Jing Xu,Samuel D Stegelmann,Charles W Jackson,Kevin B Koronowski,Kunjan R Dave,Juan I Young,Miguel A Perez-PinzonPubMed 30343466
- Autophagy 14:1976-19902018USP14 regulates DNA damage repair by targeting RNF168-dependent ubiquitination.Applications:Unspecified applicationReactive species:Unspecified reactive speciesArishya Sharma et. al.PubMed 29995557
Key facts
- Assay time
- 1h
- Sample type
- Tissue, Suspension cells, Adherent cells
- Reactive species
- Mouse, Human
What's included?
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ab221031 Histone Extraction Kit - Rapid / Ultra-pure enables the extraction of core histones from mammalian cells or tissues in 1 hour using a simple procedure with 20 minutes hands-on time.
Key facts
- Assay time
- 1h
- Sample type
- Tissue, Suspension cells, Adherent cells
- Reactive species
- Mouse, Human
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- Multi
- Appropriate long-term storage conditions
- Multi
- Storage information
- Please refer to protocols
Notes
ab221031 Histone Extraction Kit - Rapid / Ultra-pure enables the extraction of core histones from mammalian cells or tissues in 1 hour using a simple procedure with 20 minutes hands-on time. As post-translational modifications, such as acetylation, methylation and phosphorylation are maintained, histones extracted using this kit can be used in a wide variety of downstream applications.
This kit is compatible with three distinct protocols: Standard, Rapid and Ultra-pure Histone Extraction. The Standard Extraction protocol is a 1 hour procedure (of which 20 minutes is hands-on) that is suitable for histone extraction from a wide range of cell lines and tissues. The Rapid Extraction protocol can be used to prepare a crude histone extract quickly for use in applications where histone purity is not critical. It is also recommended for histone extraction using fragile cell lines or in instances where extraction of cytoplasmic histones is desirable. This protocol takes 45 minutes and requires 15 minutes of hands-on time. The Ultra-pure Histone Extraction protocol is a 75-minute procedure (with 25 minutes hands-on time) that yields very pure histones from a wide range of cell lines and tissues.
Each kit contains reagents to perform 100 extractions, where each extraction requires 2 x 106 cells or 20 mg of tissue starting material. Typical yields are around 80 μg of histones from 2 x 106 HeLa cells.
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5 product images
Functional Studies - Histone Extraction Kit - Rapid / Ultra-pure (ab221031)
Histone extraction from HeLa cells.
2 × 106 of HeLa (human epithelial cell line from cervix adenocarcinoma) cells were processed as per the standard protocol. The resulting fractions were analysed by SDS PAGE followed by Coomasie Blue staining (left panel) or Western Blot analysis (right panel). Western Blot was performed using rabbit polyclonal to H2B (Anti-Histone H2B antibody - ChIP Grade ab1790) and mouse monoclonal to H3 (Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade ab24834) primary antibodies and fluorophore-labeled secondary antibodies to assess distribution of specific histones between the fractions (right panel).
Lane 1: Pre-extract
Lane 2: Histone extract
Lane 3: Pellet
Functional Studies - Histone Extraction Kit - Rapid / Ultra-pure (ab221031)
Comparison of different histone extraction methods.
2 × 106 of HeLa (human epithelial cell line from cervix adenocarcinoma) cells were processed using competitor kit or Abcam ab221031 Histone extract kit according to standard, rapid or ultra-pure protocol. For ultra-pure protocol 176 μL pre-extraction buffer were combined with 24 μL of pre-extraction supplement. The resulting histone extract fractions were analysed by SDS PAGE.
Lane 1: Competitor kit
Lane 2: Standard protocol
Lane 3: Rapid protocol
Lane 4: Ultra-pure protocol
Functional Studies - Histone Extraction Kit - Rapid / Ultra-pure (ab221031)
Histone extraction from cell lines.
1 × 106 of indicated cells were processed as per the standard protocol. The resulting histone extract fractions were analysed by SDS PAGE followed by Coomasie Blue staining (left panel). Distribution of specific histones between pre-extract, extract and pellet fractions was measured by Western Blot using rabbit polyclonal to H2B (Anti-Histone H2B antibody - ChIP Grade ab1790) and mouse monoclonal to H3 (Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade ab24834) primary antibodies and fluorophore-labeled secondary antibodies to enable accurate signal quantification (bar charts on the right). Cell lines used in this experiment were: NIH/3T3 (mouse embryonic fibroblasts), A375 (human malignant melanoma), A673 (human Ewing's Sarcoma), SHSY-5Y (human neuroblastoma cell line).
Lane 1: NIH/3T3
Lane 2: A375
Lane 3: A673
Lane 4: SHSY-5Y
Functional Studies - Histone Extraction Kit - Rapid / Ultra-pure (ab221031)
Histone extraction from mouse kidney.
30 mg of frozen mouse kidney were processed as per the standard protocol. The resulting fractions were analysed by SDS PAGE followed by Coomasie Blue staining (left panel) or Western Blot analysis. Western Blot was performed using rabbit polyclonal to H2B (Anti-Histone H2B antibody - ChIP Grade ab1790) and mouse monoclonal to H3 (Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade ab24834) primary antibodies and fluorophore-labeled secondary antibodies to assess distribution of specific histones between the fractions (right panel).
Lane 1: Pre-extract
Lane 2: Histone extract
Lane 3: Pellet
Functional Studies - Histone Extraction Kit - Rapid / Ultra-pure (ab221031)
Histone extraction from mouse liver.
30 mg of frozen mouse liver were processed as per the standard protocol. The resulting fractions were analysed by SDS PAGE followed by Coomasie Blue staining (left panel) or Western Blot analysis. Western Blot was performed using rabbit polyclonal to H2B (Anti-Histone H2B antibody - ChIP Grade ab1790) and mouse monoclonal to H3 (Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade ab24834) primary antibodies and fluorophore-labeled secondary antibodies to assess distribution of specific histones between the fractions (right panel).
Lane 1: Pre-extract
Lane 2: Histone extract
Lane 3: Pellet
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