WB
Cell treatment (CT) Western blot analysis of Histone H3 (mutated K27 M) on different lysates with Rabbit anti-Histone H3 (mutated K27 M) antibody at 1/1,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 2 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.ICC/IF
Cell treatment (CT) Immunocytochemistry analysis of HeLa cells transfected with Histone H3 / Histone H3 (mutated K27 M) labeling Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody at 1/1,000 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 (mutated K27 M) antibody at 1/1,000 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.IHC
Cell treatment (CT) Immunohistochemical analysis of paraffin-embedded 293T cells transfected with Histone H3 / Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 1/1,000 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.CHIP
Cell treatment (CT) Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells transfected with Histone H3 (mutated K27 M) cells with Histone H3 (mutated K27 M) 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 (mutated K27 M) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human Histone H3.3 aa 1-50 (mutated K27 M). |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPICC/IFIHCWB |
| WB:1:1000-1:5000 ICC/IF:1:1000 IHC:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | H3C1 |
|---|---|
| Gene Synonyms | H3/A H3FA 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] |
| Molecular Weight(MW) | 15 kDa(Observed band size: 18 kDa) |
| Cellular Localization | Nucleus, Chromosome. |
| SwissProt ID | P68431 |
|---|
WB
Cell treatment (CT) Western blot analysis of Histone H3 (mutated K27 M) on different lysates with Rabbit anti-Histone H3 (mutated K27 M) antibody at 1/1,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 2 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.
ICC/IF
Cell treatment (CT) Immunocytochemistry analysis of HeLa cells transfected with Histone H3 / Histone H3 (mutated K27 M) labeling Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody at 1/1,000 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 (mutated K27 M) antibody at 1/1,000 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.
IHC
Cell treatment (CT) Immunohistochemical analysis of paraffin-embedded 293T cells transfected with Histone H3 / Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 1/1,000 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.
CHIP
Cell treatment (CT) Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells transfected with Histone H3 (mutated K27 M) cells with Histone H3 (mutated K27 M) 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:1:1000-1:5000 ICC/IF:1:1000 IHC:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
