重组Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27)抗体[RM1045]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/50 (9.54 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing increased nuclear staining in HeLa cells treated with 500 ng/mL TSA for 4 hours is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h (Red) / Untreated control (Green) cells labelling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/500 dilution (0.1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/2000 (0.239 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on human colon. The section was incubated with ab300641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/2000 (0.239 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on human tonsil. The section was incubated with ab300641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of paraffin-embedded Human Prostatic hype tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/2000 (0.239 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on human prostatic hyperplasia. The section was incubated with ab300641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IP

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Immunoprecipitation - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Histone H3(acetyl K4+K9+K14+K18+K23+K27) was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h whole cell lysate 10 µg with ab300641 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300641 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (ab300641) at 1/30 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) (Input) treated with 500ng/ml TSA for 4h whole cell lysate at 10 µg

Lane 2:

ab300641 IP in HeLa treated with 500ng/ml TSA for 4h whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300641 in HeLa treated with 500ng/ml TSA for 4h whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 100s

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse colon (fresh) tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/500 (0.954 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on mouse colon is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/2000 (0.239 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on rat colon. The section was incubated with ab300641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/2000 (0.239 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on mouse colon. The section was incubated with ab300641 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat colon (fresh) tissue labeling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/500 (0.954 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on rat colon is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) treated with 500ng/ml TSA for 4h (Red) / Untreated control (Green) cells labelling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/500 dilution (0.1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Histone H3(acetyl K4+K9+K14+K18+K23+K27) with ab300641 at 1/50 (9.54 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing increased nuclear staining in NIH/3T3 cells treated with 500 ng/mL TSA for 4 hours is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Histone H3(acetyl K4+K9+K14+K18+K23+K27) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) treated with 500ng/ml TSA for 4h whole cell lysate 10 µg with ab300641 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300641 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (ab300641) at 1/30 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) treated with 500ng/ml TSA for 4h whole cell lysate (Input) at 10 µg

Lane 2:

ab300641 IP in NIH/3T3 treated with 500ng/ml TSA for 4h whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300641 in NIH/3T3 treated with 500ng/ml TSA for 4h whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 100s

• WB

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Western blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

All lanes:

Western blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (ab300641) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

C6 treated with /ml TSA for 4h whole cell lysate 20ug at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

• WB

Supplier Data

Western blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

All lanes:

Western blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (ab300641) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with /ml TSA for 4h whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

• WB

Supplier Data

Western blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

All lanes:

Western blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (ab300641) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 treated with /ml TSA for 4h whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

• PepArr

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Peptide Array - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Peptide array analysis of ab300641 at 0.1ug/ml (0.1ug/ml) followed by a Goat Anti-Rabbit IgG (H+L) Fluo 647nm conjugated at 1 : 50000 dilution. All batches of ab300641 are tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).Circle area represents affinity between the antibody and a peptide : all antigen-containing peptides are displayed as red circles all other peptides as blue circles. The affinity is calculated as the area under the curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity. The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Product Protocol section.

• Dot

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Dot Blot - Anti-Histone H3 (acetyl K4+K9+K14+K18+K23+K27) antibody [RM1045] (AB300641)

Dot blot analysis of Histone H3(acetyl K4+K9+K14+K18+K23+K27) using ab300641 at 1 : 1000 dilution (0.477 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 10,000 dilution. Lane 1 : Histone H3 (acetyl K4) peptide Lane 2 : Histone H3 (acetyl K4) peptide Lane 3 : Histone H3 (acetyl K14) peptide Lane 4 : Histone H3 (acetyl K18) peptide Lane 5 : Histone H3 (acetyl K23) peptide Lane 6 : Histone H3 (acetyl K27) peptide Lane 7 : Histone H3 unmodified peptide Exposure time : 40 seconds. Blocking and diluting buffer and concentration : 5% NFDM/TBST