p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95]
cat.: HA722074
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse |
| Applications: | WB, IF-Cell, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH03-95 |
| 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: | IgG |
| Immunogen: | Recombinant protein within mouse p53 protein. |
| Positive control: | RAW264.7 cell lysate, Neuro-2a cell lysate, NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours lysate, NIH/3T3. |
| Subcellular location: | Nucleus. Cytoplasm. Localized in both nucleus and cytoplasm in most cells. In some cells, forms foci in the nucleus that are different from nucleoli. |
| Recommended Dilutions:
WB IF-Cell IP |
1:1,000 1:200 1-2μg/sample |
| Uniprot #: | SwissProt: P02340 Mouse |
| 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 protein 53 Tumor protein p53 Tumor suppressor p53 |
Images
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Fig1:
Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (HA722074) at 1/1,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: Neuro-2a cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 53 kDa Observed band size: 53 kDa Exposure time: 3 minutes 20 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 (HA722074) 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 NIH/3T3 cells treated with 0.5μM doxorubicin for 20 hours labeling p53 with Rabbit anti-p53 antibody (HA722074) at 1/200 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-p53 antibody (HA722074) at 1/200 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:
p53 was immunoprecipitated in 0.2mg NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate with HA722074 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722074 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate (input) Lane 2: HA722074 IP in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate Lane 3: Rabbit IgG instead of HA722074 in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute; ECL: K1802 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.