Application
Fig1: Western blot analysis of Tri-Methyl-Histone H3 (Lys9) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1112-3, 1/2,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane A: F9 cell lysate Lane B: F9+non-methyl peptide Lane C: F9+Tri-methl (lys9) peptide Lane D: PC12 cell lysate Lane E: NCCIT cell lysateApplication
Fig2: ICC staining of Tri-Methyl-Histone H3 (Lys9) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1112-3, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.Application
Fig3: Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Tri-Methyl-Histone H3 (Lys9) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M1112-3, 1/100) for 30 minutes 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.| Product Name | Histone H3 (tri methyl K9) [1-6] |
|---|---|
| Antibody Type | Primary Antibodies |
| Antigen Alias | 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 |
| 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 fibers. Two molecules of each of the four core histones (H2A, H2B, H3 and H4) form the octamer, which is comprised of two H2A-H2B dimers and two H3-H4 dimers, forming two nearly symmetrical halves by tertiary structure. Histones are subject to posttranslational modification by enzymes primarily on their N-terminal tails, but also in their globular domains. Human Histone H3 is subject to trimethylation at Lys 9, a modification that may be necessary for select DNA transactions or chromatin state transitions. Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me). Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription. Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). |
| Immunogen | Synthetic peptide within N terminal human Histone H3 including tri methyl K9 conjugated to keyhole limpet haemocyanin. |
| Modification | tri methyl K9 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG1 |
| Host Species | Mouse |
| Tested Applications | WBICCIHC |
| WB:1:5,000-1:10,000: ICC:1:500-1:1,000: IHC:1:200: |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| 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 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 |
|---|---|
| Molecular Weight(MW) | 17 kDa |
| Cellular Localization | Nucleus. |
Application
Fig1: Western blot analysis of Tri-Methyl-Histone H3 (Lys9) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1112-3, 1/2,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane A: F9 cell lysate Lane B: F9+non-methyl peptide Lane C: F9+Tri-methl (lys9) peptide Lane D: PC12 cell lysate Lane E: NCCIT cell lysate
Application
Fig2: ICC staining of Tri-Methyl-Histone H3 (Lys9) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1112-3, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
Application
Fig3: Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-Tri-Methyl-Histone H3 (Lys9) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M1112-3, 1/100) for 30 minutes 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.| Positive Control | NCCIT cell, F9 cell, PC12 cell, A549 cell. |
|---|---|
| Application Notes | WB:1:5,000-1:10,000: ICC:1:500-1:1,000: IHC:1:200: |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 50%Glycerol. Preservative: 0.05% Sodium Azide. |
