Mouse monoclonal antibody against P53 (Clone ISM008)
Mouse monoclonal antibody against P53 (Clone ISM008)
Product identification
ISC148-R3 3 ml ready-to-use (RTU)
ISC148-R7 7 ml ready-to-use (RTU)
ISC148-C02 0.2 ml concentrated
ISC148-C05 0.5 ml concentrated
Summary and explanation
The p53 gene is a tumour suppressor gene located on the short arm of chromosome 17 (17p13.1). The p53 (phospho-)protein (p53), 53 kDa, is expressed in the nuclei of all normal cells, but usually not immunohistochemically detectable due to a very short half-life (10-20 min.). p53 is called the guardian of the genome. By binding to DNA, the normal p53 negatively regulates cell growth and division. In case of DNA damage, p53 arrests the cell cycle until repair has taken place. In case repair is not possible, p53 induces apoptosis. p53 acts a two checkpoints in the cell cycle, between G1 and S, and between G2 and M, respectively.
Principle of the procedure
The stated primary antibody is suitable for immunohistochemical staining of FFPE tissue sections based on specific antigen-antibody reaction. Using a detection system linked to HRP or alkaline phosphatase the antigen visualization is performed via specific binding of the primary antibody. Secondary antibody is binding to the primary antibody, and the enzyme complex labels this complex. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Each step is incubated for a precise time and temperature and requires interposed washing steps. The specimen may then be counterstained. Results are interpreted using a light microscope
Staining procedure
- Cut 3-4 μm section of formalin-fixed paraffin-embedded tissue and place on positively charged slides.
- Dry at 65°C for 2 hours.
- Deparaffinize, rehydrate, and epitope retrieve. Upon completion, rinse with 3 changes of distilled or deionized water.
- If using HRP detection system, place slides in peroxide block for 10 minutes; rinse. If using AP detection system, omit this step.
- Apply the antibody and incubate for 60 minutes; rinse.
- Apply the InSituVison™ Polymer Rabbit/Mouse Detection System for 30 minutes; rinse.
- Apply ample amount of DAB or AEC chromogen and incubate; rinse.
- Dehydrate and coverslip.
Interpretation of results
The immunostaining procedure causes a colored reaction product to precipitate at the antigen sites localized by the primary antibody.
Cellular localization: Cell nucleus.
A qualified pathologist experienced in immunohistochemistry procedures must evaluate positive and negative tissue controls before interpreting patient specimens.
Positive staining intensity should be assessed within the context of any background staining of the negative reagent control.
Note: A negative result means that the antigen in question was not detected, but not that the antigen is not present in the cells/tissues tested. An antibody panel may be used to support the results in some circumstances. Additionally, the morphology of each tissue sample should be examined utilizing a hematoxylin and eosin stained section. A qualified pathologist must interpret the patient’s morphologic findings and pertinent clinical data.
Warnings and precautions
- Application only by qualified and trained personnel.
- There are no estimated health risks, if the product is used as directed. MSDS is available on request.
- Product contains sodium azide as preservative. Pure sodium azide is toxic. The concentration of sodium azide in this reagent is < 0.1 % and is not classified hazardous.
See MSDS.
- As with any product derived from biological sources, proper handling procedures should be used.
- Do not use reagents after expiration date.
- Take reasonable precautions when handling reagents. Use protective clothing and gloves.
- All hazardous materials should be disposed according to guidelines for hazardous waste disposal. Materials of human or animal origin should be handled as biohazardous materials and disposed of with proper precautions.
- Avoid microbial contamination of reagents as it may cause incorrect results.
Materials provided
| Primary antibody | P53 |
| Host | Mouse |
| Subclass | IgG |
| Clone | ISM008 |
| Species Cross-reactivity | Human. Others-not known |
| Applications | Immunohistochemistry |
| Epitope Retrieval | Heat-induced epitope retrieval |
| Ready-to-use antibody | Prediluted antibody in antibody diluent buffer |
| Recommended working dilution range | 1:50 to 1:100 |
Product label shows the specific lot number.
Prediluted antibody is ready-to-use and optimized for staining. No further dilution, reconstitution, mixing, or titration is needed.
Antibody concentrate is optimized for dilution within dilution range using InSituChem® Antibody Diluent for IHC (Cat. No. D005-50). Indicated dilution range should be considered as recommendation and depends on different facts (tissue, fixation, incubation conditions, etc.). Optimum dilution to be determined in user’s own system.
Materials required but not provided
The following materials may be required for staining but are not provided with the primary antibody.
– Positive and negative controls
– Microscope slides (positively charged) and cover slips
– Water bath
– Humidified chamber
– Staining jars
– Stopwatch
– Xylene or xylene substitute
– Ethanol
– Deionized or distilled water
– Antigen retrieval reagent, e.g. Antigen Enhancer (HIER buffer), Cat. No. D004-500
– Detection system, e.g. IHC Complete Detection system (Goat anti mouse/rabbit HRP, DAB staining), Cat. No. D001-15
– Wash buffer: e.g. IHC Wash Buffer, Cat. No. D003-500
– Tap water/bluing reagent (e.g. ammonia water)
– Light microscope
Storage and handling
Store at 2 – 8 °C.
When stored correctly antibody is stable to the expiration date indicated on the vial. Do not use after expiration date.
Literature
[1] Walerych D, Lisek K, Del Sal G. Mutant p53: One, No One, and One Hundred Thousand. Front Oncol. 2015 Dec 21;5:289.
[2] Miciak J, Bunz F. Long story short: p53 mediates innate immunity. Biochim Biophys Acta. 2016 Apr;1865(2):220-7.
[3] Nakamura Y, Arakawa H.Discovery of Mieap-regulated mitochondrial quality control as a new function of tumor suppressor p53. Cancer Sci. 2017 Feb 21.
Date of publication or revision
2024-06-10
Change(s) made:
Explanation of symbols
