Anti-p53 antibody [SP161] (ab227655)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP161] to p53
- Suitable for: Flow Cyt, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-p53 antibody [SP161]
See all p53 primary antibodies -
Description
Rabbit monoclonal [SP161] to p53 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human colon adenocarcinoma, stomach adenocarcinoma, ovarian adenocarcinoma, bladder transitional cell carcinoma, lung squamous cell carcinoma, B-cell lymphoma and breast ductal carcinoma tissue. ICC/IF: MCF7 cells. Flow Cyt: HAP1 and MCF7 cells
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General notes
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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Purity
Protein A/G purified -
Purification notes
Purified from TCS by protein A/G. -
Clonality
Monoclonal -
Clone number
SP161 -
Isotype
IgG -
Research areas
Images
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Formalin-fixed, paraffin-embedded human stomach adenocarcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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Flow cytometry overlay histogram showing wild-type HAP1 (green line) and TP53 knockout HAP1 cells stained with ab227655 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab227655) (1x106 in 100 µl at 0.008 µg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150177) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line TP53 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
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Flow Cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling p53 with purified ab227655 at 1/20 dilution (7.5 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.
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Immunocytochemistry/ Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling p53 with purified ab227655 at 1/20 (7.25µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human bladder transitional cell carcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human lung squamous cell carcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human ovarian adenocarcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human B-cell lymphoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for p53 using ab227655 at 1/100 dilution in immunohistochemical analysis.
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