Product Type: | Recombinant Rabbit monoclonal IgG1, primary antibodies |
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Species reactivity: | Human |
Applications: | WB, IF-Cell |
Clonality: | Monoclonal |
Clone number: | PSH01-98 |
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: 53 kDa |
Isotype: | IgG1 |
Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser15 of Human p53 aa 1-50 / 393. |
Positive control: | HT-29 treated with 0.5μM Doxorubicin for 24 hours cell lysate, HT-29 treated with 0.5μM Doxorubicin for 24 hours cell. |
Subcellular location: | Cytoplasm, Nucleus, Nucleus, PML body, Endoplasmic reticulum, Mitochondrion matrix, Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. |
Recommended Dilutions:
WB IF-Cell |
1:1,000 1:10,000 |
Uniprot #: | SwissProt: P04637 Human |
Alternative names: | Antigen NY-CO-13 BCC7 Cellular tumor antigen p53 FLJ92943 LFS1 Mutant tumor protein 53 p53 p53 tumor suppressor P53_HUMAN Phosphoprotein p53 Tp53 Transformation related protein 53 TRP53 tumor antigen p55 Tumor protein 53 Tumor protein p53 Tumor suppressor p53 |
Fig1:
Western blot analysis of Phospho-p53 (S15) on different lysates with Rabbit anti-Phospho-p53 (S15) antibody (HA721756) at 1/1,000 dilution. Lane 1: HT-29 cell lysate Lane 2: HT-29 treated with 0.5μM Doxorubicin for 24 hours cell lysate Lane 3: HT-29 treated with 0.5μM Doxorubicin for 24 hours, then treated with λpp for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 53 kDa Observed band size: 53 kDa Exposure time: 1 minute 46 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 (HA721756) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of HT-29 treated with or without 0.5μM Doxorubicin for 24 hours cells labeling Phospho-p53 (S15) with Rabbit anti-Phospho-p53 (S15) antibody (HA721756) at 1/10,000 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-p53 (S15) antibody (HA721756) at 1/10,000 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. |