WB
Western blot analysis of Histone H3 (acetyl K56) on different lysates with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: C6 cell lysate, Lane 3: C6 treated with 1μM TSA for 18 hours cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 14 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Histone H3 (acetyl K56) with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (acetyl K56) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | Histone H3 (acetyl K56) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Eukaryotic histones are basic and water soluble nuclear proteins that form hetero-octameric nucleosome particles by wrapping 146 base pairs of DNA in a left-handed super-helical turn sequentially to form chromosomal fiber. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form the octamer; formed of two H2A-H2B dimers and two H3-H4 dimers, forming two nearly symmetrical halves by tertiary structure. Over 80% of nucleosomes contain the linker Histone H1, derived from an intronless gene, that interacts with linker DNA between nucleosomes and mediates compaction into higher order chromatin. Histones are subject to posttranslational modification by enzymes primarily on their N-terminal tails, but also in their globular domains. Such modifications include methylation, citrullination, acetylation, phosphorylation, sumoylation, ubiquitination and ADP-ribosylation. |
| Immunogen | Synthetic peptide within Human Histone H3 aa 31-80 / 136. |
| Modification | acetyl K56 |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:500-1:2000 IHC:1:200-1:5000 ICC/IF:1:50-1:200 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | H3.3A antibody
HIST1 cluster H3E antibody H3 histone family member A antibody H3.1 antibody H3/l antibody H3F3 antibody H3FF antibody H3FJ antibody H3FL antibody Histone gene cluster 1 H3 histone family member E antibody histone H3.1t antibody Histone H3/o antibody FLJ92264 antibody H 3 antibody H3 antibody H3 histone family member B antibody H3 histone family member C antibody H3 histone family member D antibody H3 histone family member F antibody H3 histone family member H antibody H3 histone family member I antibody H3 histone family member J antibody H3 histone family member K antibody H3 histone family member L antibody H3 histone family member T antibody H3 histone family 3A antibody H3/A antibody H3/b antibody H3/c antibody H3/d antibody h3/f antibody H3/h antibody H3/i antibody H3/j antibody H3/k antibody H3/t antibody H31_HUMAN antibody H3F1K antibody H3F3A antibody H3FA antibody H3FB antibody H3FC antibody H3FD antibody H3FH antibody H3FI antibody H3FK antibody HIST1 cluster H3A antibody HIST1 cluster H3B antibody HIST1 cluster H3C antibody HIST1 cluster H3D antibody HIST1 cluster H3F antibody HIST1 cluster H3G antibody HIST1 cluster H3H antibody HIST1 cluster H3I antibody HIST1 cluster H3J antibody HIST1H3A antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody HIST1H3E antibody HIST1H3F antibody HIST1H3G antibody HIST1H3H antibody HIST1H3I antibody HIST1H3J antibody HIST3H3 antibody Histone 1 H3a antibody Histone 1 H3b antibody Histone 1 H3c antibody Histone 1 H3d antibody Histone 1 H3e antibody Histone 1 H3f antibody Histone 1 H3g antibody Histone 1 H3h antibody Histone 1 H3i antibody Histone 3 H3 antibody histone cluster 1 H3 family member a antibody histone cluster 1 H3 family member b antibody histone cluster 1 H3 family member c antibody histone cluster 1 H3 family member d antibody histone cluster 1 H3 family member e antibody histone cluster 1 H3 family member f antibody histone cluster 1 H3 family member g antibody histone cluster 1 H3 family member h antibody histone cluster 1 H3 family member i antibody histone cluster 1 H3 family member j antibody Histone cluster 1 H3a antibody Histone cluster 1 H3b antibody Histone cluster 1 H3c antibody Histone cluster 1 H3d antibody Histone cluster 1 H3e antibody Histone cluster 1 H3f antibody Histone cluster 1 H3g antibody Histone cluster 1 H3i antibody Histone cluster 1 H3j antibody Histone gene cluster 1 H3 histone family member A antibody Histone gene cluster 1 H3 histone family member B antibody Histone gene cluster 1 H3 histone family member C antibody Histone gene cluster 1 H3 histone family member D antibody Histone gene cluster 1 H3 histone family member F antibody Histone gene cluster 1 H3 histone family member G antibody Histone gene cluster 1 H3 histone family member H antibody Histone gene cluster 1 H3 histone family member I antibody Histone gene cluster 1 H3 histone family member J antibody Histone gene cluster 1 H3A antibody Histone gene cluster 1 H3B antibody Histone gene cluster 1 H3C antibody Histone gene cluster 1 H3D antibody Histone gene cluster 1 H3E antibody Histone gene cluster 1 H3F antibody Histone gene cluster 1 H3G antibody Histone gene cluster 1 H3H antibody Histone gene cluster 1 H3I antibody Histone gene cluster 1 H3J antibody Histone H 3 antibody Histone H3.1 antibody Histone H3.2 antibody Histone H3.3 antibody Histone H3/a antibody Histone H3/b antibody Histone H3/c antibody Histone H3/d antibody Histone H3/f antibody Histone H3/h antibody Histone H3/i antibody Histone H3/j antibody Histone H3/k antibody Histone H3/l antibody Histone H3/m antibody |
|---|---|
| Molecular Weight(MW) | 15 kDa |
| Cellular Localization | Nucleus, Chromosome, Nucleosome core. |
WB
Western blot analysis of Histone H3 (acetyl K56) on different lysates with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: C6 cell lysate, Lane 3: C6 treated with 1μM TSA for 18 hours cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 14 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Histone H3 (acetyl K56) with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (acetyl K56) antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (acetyl K56) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:500-1:2000 IHC:1:200-1:5000 ICC/IF:1:50-1:200 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
