Anti-p53 antibody [SP5] (ab16665)
Key features and details
- Rabbit monoclonal [SP5] to p53
- Suitable for: IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-p53 antibody [SP5]
See all p53 primary antibodies -
Description
Rabbit monoclonal [SP5] 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 within Human p53. The exact sequence is proprietary.
Database link: P04637 -
Epitope
Not determined -
Positive control
- IHC-P: Human colon adenocarcinoma, lung adenocarcinoma, stomach adenocarcinoma, ovarian adenocarcinoma, bladder transitional cell carcinoma, hepatocellular cell carcinoma, cervical squamous cell carcinoma, breast ductal cell carcinoma and B cell lymphoma tissue. Flow Cyt: HAP1 and MCF7 cells. ICC/IF: MCF7 cells. ICC/IF KO: Hap1 cells (Hap1-TP53 KO used as a negative cell line)
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General notes
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
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: 7.50
Preservative: 0.1% Sodium azide
Constituents: Tissue culture supernatant, Tris buffered saline, 1% BSA -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
SP5 -
Isotype
IgG -
Research areas
Images
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Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for p53 using ab16665 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 ab16665 (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 (ab16665) (1x106 in 100 µl at 0.008 µg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) 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) labeling p53 with purified ab16665 at 1/200 dilution (0.81 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730)(black). Unlableled control - Unlabelled cells (blue).
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ab16665 staining p53 in wild-type Hap1 cells (top panel) and p53 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab16665 at 1/25 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).
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Immunocytochemistry/ Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling p53 with purified ab16665 at 1/25 (6.5 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. 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 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 lung adenocarcinoma tissue stained for p53 using ab16665 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human stomach adenocarcinoma tissue stained for p53 using ab16665 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human ovarian adenocarcinoma tissue stained for p53 using ab16665 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 ab16665 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human hepatocellular cell carcinoma tissue stained for p53 using ab16665 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human cervical squamous cell carcinoma tissue stained for p53 using ab16665 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human breast ductal cell carcinoma tissue stained for p53 using ab16665 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human B cell lymphoma tissue stained for p53 using ab16665 at 1/100 dilution in immunohistochemical analysis.