p53 Antibody (BP53-12)
Novus Biologicals | Catalog # NBP2-29453
Key Product Details
Species Reactivity
Validated:
Human, Canine, Chicken, Hamster, Monkey, Mouse (Negative), Rat (Negative)
Cited:
Human, Mouse, Avian - Chicken, Canine
Applications
Validated:
Multiplex Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Simple Western, COMET
Cited:
Western Blot, Simple Western
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2a Kappa Clone # BP53-12
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Product Specifications
Immunogen
Recombinant human wild-type p53 protein (Uniprot: P04637)
Epitope
aa 16-25
Reactivity Notes
Does not react with Mouse or Rat.
Localization
Predominantly nuclear
Specificity
This monoclonal antibody reacts with an N-terminal epitope (aa 16-25) of both wild type and mutated p53. Mutation and/or allelic loss of p53 is one of the causes of a variety of mesenchymal and epithelial tumors. If it occurs in the germ line, such tumors run in families. In most transformed and tumor cells the concentration of p53 is increased 51000 fold over the minute concentrations (1000 molecules cell) in normal cells, principally due to the increased half-life (4 h) compared to that of the wild-type (20 min). p53 Localizes in the nucleus, but is detectable at the plasma membrane during mitosis and when certain mutations modulate cytoplasmic/nuclear distribution. Mutations arise with an average frequency of 70% but incidence varies from zero in carcinoid lung tumors to 97% in primary melanomas. High concentrations of p53 protein are transiently expressed in human epidermis and superficial dermal fibroblasts following mild ultraviolet irradiation. Positive nuclear staining with p53 antibody has been reported to be a negative prognostic factor in breast carcinoma, lung carcinoma, colorectal, and urothelial carcinoma. Anti-p53 positivity has also been used to differentiate uterine serous carcinoma from endometrioid carcinoma as well as to detect intratubular germ cell neoplasia.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2a Kappa
Theoretical MW
53 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Description
200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-33074)
Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.
Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.
Scientific Data Images for p53 Antibody (BP53-12)
Detection of p53 in Human Colon Tumor via seqIF™ staining on COMET™
p53 was detected in immersion fixed paraffin-embedded sections of human Colon Tumor using Mouse Anti-Human p53 Monoclonal Antibody (Catalog # NBP2-29453) at 12ug/mL at 37 ° Celsius for 8 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Mouse IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the astrocytes showing nuclear staining. Protocol available in COMET™ Panel Builder.Western Blot: p53 Antibody (BP53-12) [NBP2-29453]
Western Blot: p53 Antibody (BP53-12) [NBP2-29453] - Western Blot Analysis of human HeLa cell lysate using p53 Antibody (BP53-12).Immunohistochemistry-Paraffin: p53 Antibody (BP53-12) [NBP2-29453]
Immunohistochemistry-Paraffin: p53 Antibody (BP53-12) [NBP2-29453] - p53 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using a 1 ug/mL mouse anti-p53 monoclonal (NBP2-29453, Novus Biologicals) for 1 hour at room temperature followed by the appropriate HRP polymer reagent, anti-mouse IgG VisUCyte HRP polymer (VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue).Western Blot: p53 Antibody (BP53-12) [NBP2-29453]
Western Blot: p53 Antibody (BP53-12) [NBP2-29453] - Analysis of p53 1) A431 2) MCF7 and 3) HEK293 lysate probed with p53 antibody at 1 ug/ml. goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used for this test.Immunohistochemistry-Paraffin: p53 Antibody (BP53-12) [NBP2-29453]
Immunohistochemistry-Paraffin: p53 Antibody (BP53-12) [NBP2-29453] - Formalin-paraffin human colon carcinoma stained with p53 Ab (BP53-12).Simple Western: p53 Antibody (BP53-12) [NBP2-29453]
Simple Western: p53 Antibody (BP53-12) [NBP2-29453] - Simple Western lane view shows a specific band for p53 in 0.2 mg/ml of MDA-MB-231 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.Simple Western: p53 Antibody (BP53-12) [NBP2-29453]
Simple Western: p53 Antibody (BP53-12) [NBP2-29453] - Electropherogram image of the corresponding Simple western lane view. p53 antibody was used at 10 ug/ml dilution on MDA-MB-231 lysate(s) respectively.Applications for p53 Antibody (BP53-12)
Application
Recommended Usage
Immunohistochemistry-Paraffin
1-2 ug/ml
Multiplex Immunofluorescence
12ug/mL
Simple Western
10 ug/ml
Western Blot
1-2 ug/ml
Application Notes
Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
Optimal dilution for a specific application should be determined.
In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in MDA-MB-231 lysate, separated by Size, antibody dilution of 10 ug/mL, apparent MW was 60 kDa.
Optimal dilution for a specific application should be determined.
In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in MDA-MB-231 lysate, separated by Size, antibody dilution of 10 ug/mL, apparent MW was 60 kDa.
Reviewed Applications
Read 1 review rated 5 using NBP2-29453 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified
Formulation
10 mM PBS with 0.05% BSA
Preservative
0.05% Sodium Azide
Concentration
0.2 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C.
Background: p53
Additional p53 Products
Product Documents for p53 Antibody (BP53-12)
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Product Specific Notices for p53 Antibody (BP53-12)
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Citations for p53 Antibody (BP53-12)
Customer Reviews for p53 Antibody (BP53-12) (1)
5 out of 5
1 Customer Rating
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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