Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
---|---|
Species reactivity: | Human, Mouse, Rat, Monkey |
Applications: | WB, IF-Cell, ChIP, Dot Blot, IHC-P |
Clonality: | Monoclonal |
Clone number: | JE00-98 |
Form: | Liquid |
Storage condition: | 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. |
Concentration: | 1ug/ul |
Purification: | Protein A affinity purified. |
Molecular weight: | Predicted band size: 15 kDa |
Isotype: | IgG |
Immunogen: | Synthetic peptide corresponding to the amino terminus of histone H3 in which Lys4 is di-methylated. |
Positive control: | HeLa cell lysate, MCF7 cell lysate, HCT 116 cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HeLa, NIH/3T3, PC-12, mouse colon tissue, rat colon tissue. |
Subcellular location: | Nucleus, Chromosome. |
Recommended Dilutions:
WB IF-Cell ChIP Dot Blot IHC-P |
1:1,000 1:500-1:2,000 Use 0.5~2 μg for 25 μg of chromatin. 1:1,000 10,000 |
Uniprot #: | SwissProt: P68431 human | P84243 human | Q16695 human | Q6NXT2 human | Q71DI3 human | P68433 mouse | P84228 mouse | Q6LED0 rat |
Alternative names: | H3 histone family, member A H3/A H31_HUMAN H3FA Hist1h3a HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J histone 1, H3a Histone cluster 1, H3a Histone H3.1 Histone H3/a Histone H3/b Histone H3/c Histone H3/d Histone H3/f Histone H3/h Histone H3/i Histone H3/j Histone H3/k Histone H3/l H3K4Ac |
![]() |
Fig1:
Western blot analysis of Histone H3 (di methyl K4) on different lysates with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: HCT 116 cell lysate Lane 4: COS-1 cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: C6 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 14 seconds; ECL: K1801; 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 (HA722135) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
![]() |
Fig2:
Immunocytochemistry analysis of HeLa cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) at 1/500 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 (di methyl K4) antibody (HA722135) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
![]() |
Fig3:
Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) 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 (di methyl K4) antibody (HA722135) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
![]() |
Fig4:
Immunocytochemistry analysis of PC-12 cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) at 1/2,000 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 (di methyl K4) antibody (HA722135) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
![]() |
Fig5: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H3 (di methyl K4) (HA722135) 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. |
![]() |
Fig6:
Dot blot analysis of Histone H3 (di methyl K4) on different proteins with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature. Lane 1: Unmodified Histone H3 (negative) Lane 2: Mono-Methyl-Histone H3 (Lys4) (negative) Lane 3: Di-Methyl-Histone H3 (Lys4) (positive) Lane 4: Tri-Methyl-Histone H3 (Lys4) (negative) Lane 5: Acetyl-Histone H3 (Lys4) (negative) Lane 6: Acetyl-Histone H3 (Lys9) (negative) Proteins loading: 100ng, 25ng, 5ng, 1ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 15 seconds; ECL: K1801. |
![]() |
Fig7:
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) at 1/10,000 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 (HA722135) at 1/10,000 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. |
![]() |
Fig8:
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Histone H3 (di methyl K4) antibody (HA722135) at 1/10,000 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 (HA722135) at 1/10,000 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. |