Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade

14 product images

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  ChIP - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Chromatin from Xenopus laevis oocytes was prepared according to the Abcam X-ChIP protocol.
  

  Oocytes were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 mg of chromatin, 3 mg of ab7834 (anti-H3, light blue) and 3 μg of ab1791 (anti-H3, dark blue), and 20 ml of Protein A/G sepharose beads. A non-specific antibody was used as a control (yellow).

  The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).

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  ChIP - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  The ChIP was performed with chromatin from mouse gut cell lysate and ab1791 at 1/250 dilution.
  

  Negative control: No antibody was used (right bar).

  The immunoprecipitated DNA was quantified by real time PCR.

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  Immunoprecipitation - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Histone H3 - ChIP Grade was immunoprecipitated using 0.5mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extract, 5 μg of Rabbit polyclonal to and 50 μl of protein G magnetic beads (+). No antibody was added to the control (-).
  

  The antibody was incubated under agitation with Protein G beads for 10 minutes, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 minutes under agitation.

  Proteins were eluted by addition of 40 μl SDS loading buffer and incubated for 10 minutes at 70°C; 10 μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab1791.

  Secondary Antibody: Mouse anti-rabbit HRP light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).

  Band: 15kDa; Histone H3 - ChIP Grade

  All lanes: Immunoprecipitation - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 15 kDa

  Exposure time: 30s

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  Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab1791 overnight at 4°C.
  

  Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody was used for detection.

  Antibody binding was visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1/1000 dilution

  Lanes 1 and 4: A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

  Lanes 2 and 5: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 20 µg

  Lanes 3 and 6: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 15 kDa

  Observed band size: 17 kDa

  Exposure time: 10s

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  This image is courtesy of an anonymous customer review

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Histone H3 immunohistochemistry staining of mouse liver using rabbit anti-Histone H3 antibody
  
  ab1796 staining Histone H3 in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed paraffin-embedded sections).

  Tissue was fixed with paraformaldehyde permeabilized with 0.05% Triton X-100 in PBS for 30 minutes and blocked with 5% BSA for 1 hour; antigen retrieval was by heat mediation in sodium citrate pH 6. Samples were incubated with the primary antibody (1/500 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG polyclonal (1/400 dilution) was used as the secondary antibody.

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  ab1791 staining Histone H3 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab1791 at 0.1 μg/mL and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

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  This image is courtesy of an anonymous customer review

  Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Blocked with 3% milk for 1 hour at 25°C.
  

  Incubated with the primary antibody for 16 hours at 4°C in 3% milk in TBS-tween.

  All lanes: Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1/1000 dilution

  Lane 1: Mouse skeletal muscle mitochondrial fraction at 20 µg

  Lane 2: Mouse skeletal muscle nuclear fraction at 20 µg

  Secondary

  All lanes: HRP-conjugated goat anti-rabbit IgG at 1/4000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 15 kDa

  Observed band size: 17 kDa

  Exposure time: 7min

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  This image is courtesy of an anonymous customer review

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  ab1791 staining Histone H3 in HeLa (Human epithelial cell line from cervix adenocarcinoma) by ICC/IF (Immunocytochemistry/immunofluorescence).
  

  Cells were fixed with methanol and blocked with 0.2% fish scale gelatin for 1 hour at 25°C. Samples were incubated with the primary antibody (1/300 in PBS + 0.2% gelatin) for 20 minutes at 25°C. An Alexa Fluor^® 488-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

  Green - Histone H3.
  Blue - DAPI.
  Red - Tubulin.

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  Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  ab1791 is tested in western blot on a range of species. We recommend loading higher amounts of protein (20-30ug) to increase the signal in yeast lysates

  All lanes: Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 2: Western blot - NIH/3T3 whole cell lysate (NIH/3T3 whole cell lysate ab7179) at 10 µg

  Lane 3: Drosophila embryo nuclear extract (from melanogaster embryos 0-12Hr) at 10 µg

  Lane 4: S.cerevisiae (Y190) Whole Cell Lysate at 10 µg

  Lane 5: S.pombe Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

  Performed under reducing conditions.

  Predicted band size: 15 kDa

  Observed band size: 17 kDa

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  This image is courtesy of John E. Mueller and J. Ruth German (Mary Bryk lab)

  Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Rabbit polyclonal to Histone H3 (ab1791) at 1/5000 on S. cerevisiae whole cell lysate (40 ug per lane).
  
  Protein resolved on 15% SDS-PAGE gel. After transfer to PVDF membrane, blots were blocked in 1X PBS, 0.1% Tween-20, and 5% milk. ab1791 was diluted in 5 ml blocking buffer at 1/5000. Blots plus primary antibodies were either incubated overnight at 4°C or at RT for 2 hours. Blots were washed 6X for 10 minutes each in PBS with 0.1% Tween-20 before addition of secondary antibodies. Secondary antibodies were diluted 1/2,000 in blocking buffer and incubated with blots for 2 hours at RT. Secondary blots were washed 4X for 10 minutes each in PBS with 0.1% Tween-20 and 2X for 10 minutes each in PBS.

  All lanes: Western blot - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Predicted band size: 15 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Histone H3 immunohistochemistry staining of rat brain using rabbit anti-Histone H3 antibody
  
  Paraffin-embedded rat brain tissue stained for Histone H3 using ab1791 at 1/8000 dilution in immunohistochemical analysis.

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  This image is courtesy of an anonymous customer review

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  ab1791 staining Histone H3 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
  

  Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An un-diluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Histone H3 immunohistochemistry staining of rat brain using rabbit anti-Histone H3 antibody
  
  ab1791 staining Histone H3 (red) in rat brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed paraffin-embedded sections).

  Tissue was fixed with formaldehyde permeabilized with 0.1% TBS-TritonX and blocked with 10% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with the primary antibody (1/500 in 10% normal goat serum) for 24 hours at 24°C. An Alexa Fluor^® 594-conjugated goat anti-rabbit IgG polyclonal (1/500 dilution) was used as the secondary antibody.

  Green - Nucleus staining.
  Red - Histone H3 staining.

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  ChIP - Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791)

  Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab1791 (blue), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.