Phospho-Histone H3.3 (S31) Recombinant Rabbit Monoclonal Antibody [JE62-92]
cat.: HA721034
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Rat |
| Applications: | WB, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | JE62-92 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% 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 phospho-peptide corresponding to residues surrounding Ser31 of Human Histone H3.3. |
| Positive control: | HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate, PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, HeLa cells treated with 100ng/mL Nocodazole for 18 hours. |
| Subcellular location: | Nucleus, Chromosome. |
| Recommended Dilutions:
WB IF-Cell |
1:500-1:1,000 1:25,000 |
| Uniprot #: | SwissProt: P84243 Human | P84245 Rat |
| Alternative names: | H3 histone family 3A H3 histone family 3B H3 histone, family 3B (H3.3B) H3.3 H3.3A H3.3B H33_HUMAN H3F3 H3F3A H3f3b Histone H3.3 Histone H3.3Q Histone H3.A Histone H3.B MGC87782 MGC87783 |
Images
|
Fig1:
Western blot analysis of Histone H3.3 (phospho S31) on Hela cell lysates. Lane 1: Hela cells, whole cell lysate, 10ug/lane Lane 2/3: Hela cells treated with 100ng/ml Nocodazole for 18h, whole cell lysates, 10ug/lane Lane 4: Hela cells treated with 100ng/ml Nocodazole for 18h, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane All lanes : Anti-Histone H3.3 (phospho S31) antibody (HA721034) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution. Predicted band size: 15 kDa Observed band size: 15 kDa Blocking and diluting buffer: 5% BSA. Exposure time: 2 minutes 34 seconds Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721034) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Western blot analysis of Phospho-Histone H3.3 (S31) on different lysates with Rabbit anti-Phospho-Histone H3.3 (S31) antibody (HA721034) at 1/1,000 dilution. Lane 1: PC-12 cell lysate (20 µg/Lane) Lane 2: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate (20 µg/Lane) Predicted band size: 15 kDa Observed band size: 15 kDa 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 (HA721034) 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. |
|
Fig3:
Immunocytochemistry analysis of HeLa cells treated with or without 100ng/mL Nocodazole for 18 hours labeling Phospho-Histone H3.3 (S31) with Rabbit anti-Phospho-Histone H3.3 (S31) antibody (HA721034) at 1/25,000 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-Phospho-Histone H3.3 (S31) antibody (HA721034) at 1/25,000 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.