WB
Western blot analysis of Histone H3 (tri methyl K36) on different lysates with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: COS-1 cell lysate (20 µg/Lane), Lane 3: RAW264.7 cell lysate (20 µg/Lane), Lane 4: C6 cell lysate (20 µg/Lane), Lane 5: PC-12 cell lysate (20 µg/Lane), Exposure time: 1 minute; 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 1/5,000 dilution was used in primary antibody dilution 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 colon cancer tissue with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 RAW264.7 cells labeling Histone H3 (tri methyl K36) with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/100 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 (tri methyl K36) antibody at 1/100 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.DB
Dot blot analysis of Histone H3 (tri methyl K36) on different proteins with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/5,000 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 (Lys36) (negative), Lane 3: Di-Methyl-Histone H3 (Lys36) (negative), Lane 4: Tri-Methyl-Histone H3 (Lys36) (positive), Lane 5: Tri-Methyl-Histone H3 (Lys27) (negative), Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 1 minute 59 seconds.IP
Histone H3 (tri methyl K36) was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-Histone H3 (tri methyl K36) antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/20,000 dilution. HRP Conjugated 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: Rabbit anti-Histone H3 (tri methyl K36) antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Histone H3 (tri methyl K36) antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 59 seconds.ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (tri methyl K36) 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 (tri methyl K36) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide corresponding to residues surrounding tri-methyl Lys36 of human histone H3 protein. |
| Modification | tri methyl K36 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPDBICC/IFIHCIPWB |
| WB:1:5000 IHC:1:2000 ICC/IF:1:100 DB:1:5000 IP:1-2μg/sample ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | H3 histone family member E pseudogene antibody H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3F3 antibody H3FA antibody Hist1h3a antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody H3 histone family member E pseudogene antibody H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3F3 antibody H3FA 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 cluster 1 H3a antibody Histone H3 3 pseudogene 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 H3K36me3 |
|---|---|
| Molecular Weight(MW) | 15kDa |
| Function | In eukaryotes, DNA is wrapped around histone octamers to form the basic unit of chromatin structure. The octamer is composed of histones H2A, H2B, H3 and H4, and it associates with approximately 200 base pairs of DNA to form the nucleosome. The association of DNA with histones results in dense packing of chromatin, which restricts proteins involved in gene transcription from binding to DNA. p300 preferentially acetylates Histone H3 at lysines 14 and 18 and Histone H4 at lysines 5 and 8. PCAF in its native form, primarily acetylates Histone H3 at lysine 14 to a monoacetylated form, and less efficiently acetylates Histone H4 at lysine 8. Histone H4 may also be acetylated at lysines 12 and 16, and the involvement of acetylated H4 with Histones H2A, H2B and H3 suggests that acetylated histones may be involved in dynamic chromatin remodeling. |
| Cellular Localization | Nucleus, Chromosome. |
WB
Western blot analysis of Histone H3 (tri methyl K36) on different lysates with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: COS-1 cell lysate (20 µg/Lane), Lane 3: RAW264.7 cell lysate (20 µg/Lane), Lane 4: C6 cell lysate (20 µg/Lane), Lane 5: PC-12 cell lysate (20 µg/Lane), Exposure time: 1 minute; 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 1/5,000 dilution was used in primary antibody dilution 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 colon cancer tissue with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 RAW264.7 cells labeling Histone H3 (tri methyl K36) with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/100 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 (tri methyl K36) antibody at 1/100 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.
DB
Dot blot analysis of Histone H3 (tri methyl K36) on different proteins with Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/5,000 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 (Lys36) (negative), Lane 3: Di-Methyl-Histone H3 (Lys36) (negative), Lane 4: Tri-Methyl-Histone H3 (Lys36) (positive), Lane 5: Tri-Methyl-Histone H3 (Lys27) (negative), Proteins loading: 100ng, 25ng, 5ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 1 minute 59 seconds.
IP
Histone H3 (tri methyl K36) was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-Histone H3 (tri methyl K36) antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Histone H3 (tri methyl K36) antibody at 1/20,000 dilution. HRP Conjugated 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: Rabbit anti-Histone H3 (tri methyl K36) antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Histone H3 (tri methyl K36) antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 59 seconds.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (tri methyl K36) 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:1:5000 IHC:1:2000 ICC/IF:1:100 DB:1:5000 IP:1-2μg/sample ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| 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. |
