Histone Extraction Kit - Rapid / Ultra-pure

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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 (ab1790) and mouse monoclonal to H3 (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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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 (ab1790) and mouse monoclonal to H3 (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

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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

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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 (ab1790) and mouse monoclonal to H3 (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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Lab

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 (ab1790) and mouse monoclonal to H3 (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