WB
Western blot analysis of Histone H3 (acetyl K9) on different lysates with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 500ng/mL TSA for 4 hours cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate Lane 5: C6 cell lysate Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 6 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 temperatureIHC
Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/2,000 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 1/2,000 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.ICC/IF
Immunocytochemistry analysis of C6 cells labeling Histone H3 (acetyl K9) with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/10,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 K9) antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells labeling Histone H3 (acetyl K9). Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).Dot
Dot blot analysis of Histone H3 (acetyl K9) on different proteins with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution for 1 hour at room temperature. Lane 1: Unmodified Histone H3 (negative) Lane 2: Mono-Methyl-Histone H3 (Lys9) (negative) Lane 3: Tri-Methyl-Histone H3 (Lys9) (negative) Lane 4: Acetyl-Histone H3 (Lys9) (positive) Lane 5: Acetyl-Histone H3 (Lys4) (negative) Lane 6: Acetyl-Histone H3 (Lys27) (negative) Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 30 seconds;IP
Histone H3 (acetyl K9) was immunoprecipitated from 0.2 mg HeLa cell lysate with Histone H3 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Histone H3 antibody at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: Histone H3 antibody IP in HeLa cell lysate Lane 3: Rabbit IgG instead of Histone H3 antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 18 secondsChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (acetyl K9) / Competitor's antibody / 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 K9) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human Histone H3 (acetyl K9) aa 1-50. |
| Modification | acetyl K9 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPDotFCICC/IFIHCIPWB |
| WB:1:1000 IHC:1:2000 ICC/IF:1:5000-1:15000 FC:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. Dot Blot:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | H3C1 |
|---|---|
| Gene Synonyms | H3/A H3C2 H3C3 H3C4 H3C6 H3C7 H3C8 H3FA H3C10 H3C11 H3C12 HIST1H3A |
| Gene Full Name | H3 clustered histone 1 |
| Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq, Aug 2015] |
| Alternative Names | H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3FA antibody Hist1h3a antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody HIST1H3E antibody HIST1H3F antibody H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3FA antibody Hist1h3a antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody HIST1H3E antibody HIST1H3F antibody HIST1H3G antibody HIST1H3H antibody HIST1H3I antibody HIST1H3J antibody histone 1 H3a antibody Histone cluster 1 H3a antibody Histone H3.1 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 H3K9Ac |
| Molecular Weight(MW) | 15kDa |
| Cellular Localization | Nucleus, Chromosome. |
WB
Western blot analysis of Histone H3 (acetyl K9) on different lysates with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 500ng/mL TSA for 4 hours cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate Lane 5: C6 cell lysate Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 6 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 stomach tissue with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/2,000 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 1/2,000 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.
ICC/IF
Immunocytochemistry analysis of C6 cells labeling Histone H3 (acetyl K9) with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/10,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 K9) antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling Histone H3 (acetyl K9). Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Dot
Dot blot analysis of Histone H3 (acetyl K9) on different proteins with Rabbit anti-Histone H3 (acetyl K9) antibody at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution for 1 hour at room temperature. Lane 1: Unmodified Histone H3 (negative) Lane 2: Mono-Methyl-Histone H3 (Lys9) (negative) Lane 3: Tri-Methyl-Histone H3 (Lys9) (negative) Lane 4: Acetyl-Histone H3 (Lys9) (positive) Lane 5: Acetyl-Histone H3 (Lys4) (negative) Lane 6: Acetyl-Histone H3 (Lys27) (negative) Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 30 seconds;
IP
Histone H3 (acetyl K9) was immunoprecipitated from 0.2 mg HeLa cell lysate with Histone H3 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Histone H3 antibody at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: Histone H3 antibody IP in HeLa cell lysate Lane 3: Rabbit IgG instead of Histone H3 antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 18 seconds
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (acetyl K9) / Competitor's antibody / 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:1:1000 IHC:1:2000 ICC/IF:1:5000-1:15000 FC:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. Dot Blot:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
