p53 Recombinant Rabbit Monoclonal Antibody [SY010-6]
cat.: ET1605-16
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, IP, FC |
| Clonality: | Monoclonal |
| Clone number: | SY010-6 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 53 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within N-terminal human p53. |
| Positive control: | HT-29 cell lysate, A431 cell lysate, MDA-MB-231 cell lysate, HEK-293 cell lysate, MDA-MB-468 cell lysate, A431, human colon cancer tissue. |
| Subcellular location: | Cytoplasm, Nucleus, Endoplasmic reticulum, Mitochondrion matrix. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P FC |
1:1,000-1:5,000 1:100 1:50-1:200 1:1,000 1:1,000 |
| 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 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 (ET1605-16) at 1/5,000 dilution. Lane 1: HT-29 cell lysate Lane 2: A431 cell lysate Lane 3: Saos-2 cell lysate (negative) Lysates/proteins at 15 µg/Lane. Predicted band size: 53 kDa Observed band size: 53 kDa Exposure time: 5 minutes; 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 (ET1605-16) at 1/5,000 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. |
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Fig2:
Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (ET1605-16) at 1/1,000 dilution. Lane 1: MDA-MB-231 cell lysate Lane 2: HEK-293 cell lysate Lane 3: MDA-MB-468 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 53 kDa Observed band size: 53 kDa Exposure time: 1 minute; 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 (ET1605-16) 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. |
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Fig3:
Immunocytochemistry analysis of A431 cells labeling p53 with Rabbit anti-p53 antibody (ET1605-16) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 (ET1605-16) at 1/100 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 (HA601187, 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-p53 antibody (ET1605-16) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1605-16) at 1/1,000 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. |
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Fig5:
Flow cytometric analysis of A431 cells labeling p53. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1605-16, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.