Product datasheet and instructions for use

Product Datasheet and Instructions for Use
Product Code:
MP-323-CM01 (0.1ml conc)
MP-323-CM05 (0.5ml conc)
Product Description:
CD24
Concentrated Monoclonal Antibody
Control Number: 901-323-052510
ISO 9001:2000 CERTIFIED
Summary and Explanation:
Antigen detection, in tissues and cells, is a multi-step immunohistochemical process. The
initial step binds the primary antibody to its specific epitope. After labelling the antigen with a
primary antibody, a universal, affinity-purified, secondary antibody is added to bind to the
primary antibody. An enzyme label is then added to bind to the secondary antibody; this
detection of the bound antibody is evidenced by a colorimetric reaction.
CD24 (30-70kDa molecular weight) is a two chain glycosylphosphatidylinositol (GPI) -anchored glycoprotein. It is a mucin-like adhesion molecule. CD24 functionally enhances themetastatic potential of malignant-cells, as it has been also identified as a ligand of P-selectin,an adhesion receptor on activated endothelial cells and platelets. Research has shown CD24expression as a potentially significant parameter for a wide variety of human cancer diagnosisand for patient prognosis. Elevated CD24 membranous expression and, in particular,cytoplasmic staining seem to predict malignant transformation. Recent studies have shownCD24 as a prognostic marker for breast cancer and more specifically for tamoxifen-resistantbreast cancer cases. Further studies have shown a sub-population (CD44+/CD24-) of breastcancer cells reported to have stem/progenitor cell properties.
Intended Use: For In Vitro Diagnostic Use
Source: Mouse Monoclonal
Species Reactivity: Human; others not tested
Clone: SN3b
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Isotype: IgM/kappa
Total Protein Concentration: ~10 mg/ml. Call for lot specific Ig Concentration.
Epitope/Antigen: CD24
Cellular Localization: Cytoplasmic and/or cell membrane
Positive Control: Some breast cancers
Normal Tissue: Skin and kidney
Abnormal Tissue: Breast, ovarian or prostate cancer
Known Applications: Immunohistochemistry (formalin-fixed paraffin-embedded tissues)
Supplied As: Buffer with protein carrier and preservative.
Storage and Stability:
Store at 2ºC to 8ºC. Do not use after expiration date printed on vial. If reagents are stored
under conditions other than those specified in the package insert, they must be verified by the
user. Diluted reagents should be used promptly; any remaining reagent should be stored at 2ºC
to 8ºC.
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Instructions for Use:
Endogenous Peroxidase Block
Block all endogenous peroxidase activity by incubating the sections for 5 minutes with the
MenaPath Peroxide Block. Rinse slides in water, and then rinse well in buffer.
Pretreatment Protocol:
Retrieve sections with MenaPath Access Supreme or Access Revelation Solution using the
MenaPath Access Retrieval Unit at 80°C for 30 minutes followed by a wash in distilled water.
Alternatively, steam tissue sections for 45-60 minutes. Allow solution to cool for 20 minutesthen wash in distilled water.
Protein Block
Incubate sections for 10-15 minutes at room temperature with the MenaPath Background
Blocker with Casein
Primary Antibody
Dilute antibody 1:100-1:200 with MenaPath Sensitivity Enhancing Antibody Diluent B.
Incubate sections for 30 minutes at room temperature. Rinse slides x3 with buffer.
Universal Probe
Incubate sections for 10-20 minutes at room temperature with the MenaPath X-Cell Plus
Universal Probe. Rinse slides x3 with buffer
HRP Polymer
Incubate sections for 10-25 minutes at room temperature with the MenaPath X-Cell Plus HRP
Polymer. Rinse slides x3 in buffer.
Chromogen
Incubate sections for 5 minutes at room temperature with MenaPath X-Cell Plus Liquid Stable
DAB. Rinse x3 with buffer.
Counterstain:
Incubate for 30-60 seconds with MenaPath Haematoxylin. Rinse with deionized water. Apply
Bluing solution for 1 minute. Dehydrate, Clear and Mount
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Technical Note: Use TBS buffer for wash steps.
Performance Characteristics: The optimum antibody dilution and protocols for a specific
application can vary. These include, but are not limited to: fixation, heat-retrieval method,
incubation times, tissue section thickness and detection kit used. Due to the superior
sensitivity of these unique reagents, the recommended incubation times and titers listed are not
applicable to other detection systems, as results may vary. The data sheet recommendations
and protocols are based on exclusive use of MenaPath products. Ultimately, it is the
responsibility of the investigator to determine optimal conditions. These products are tools that
can be used for interpretation of morphological findings in conjunction with other diagnostic
tests and pertinent clinical data by a qualified pathologist.
Quality Control: Refer to NCCLS Quality Assurance for Immunocytochemistry approved
guidelines, December 1999 MM4-A Vol.19 No.26 for more information about Tissue
Controls.
Precautions: This antibody contains less than 0.1% sodium azide. Concentrations less than
0.1% are not reportable hazardous materials according to U.S. 29 CFR 1910.1200, OSHA
Hazard communication and EC Directive 91/155/EC. Sodium azide (NaN3) used as a
preservative is toxic if ingested. Sodium azide may react with lead and copper plumbing to
form highly explosive metal azides. Upon disposal, flush with large volumes of water to
prevent azide build-up in plumbing. (Center for disease control, 1976, National Institute of
Occupational Safety and Health, 1976)
Specimens, before and after fixation and all materials exposed to them, should be handled as if
capable of transmitting infection and disposed of with proper precautions. Never pipette
reagents by mouth and avoid contacting the skin and mucous membranes
with reagents and specimens. If reagents or specimens come in contact with sensitive areas,
wash with copious amounts of water.
Microbial contamination of reagents may result in an increase in nonspecific staining.
Incubation times or temperatures other than those specified may give erroneous results. The
user must validate any such change. The MSDS is available upon request.
Troubleshooting: Follow the antibody specific protocol recommendations according to data
sheet provided. If atypical results occur, contact Menarini’s Technical Service Helpline: on
01189 444130.
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Limitations and Warranty
There are no warranties, expressed or implied, which extend beyond this description. Menarini
is not liable for damages of any kind including: personal injury, or economic loss caused by
this product.
Reference:
1. Surowiak P et al. CD24 expression is specific for tamoxifen-resistant ductal breast cancer
cases. Anticancer Res. 2006 Jan-Feb;26(1B):629-34.
2. Baumann P et al. CD24 expression causes the acquisition of multiple cellular properties
associated with tumor growth and metastasis. Cancer Res. 2005 Dec 1;65 (23):10783-93.
3. Dontu G, Liu S, Wicha MS. Stem cells in mammary development and carcinogenisis:
implications for prevention and treatment. Stem Cell Rev. 2005;1 (3):207-13.
4. Center for Disease Control Manual. Guide: Safety Management, NO. CDC-22, Atlanta,
GA. April 30, 1976 "Decontamination of Laboratory Sink Drains to Remove Azide Salts."
5. National Committee for Clinical Laboratory Standards (NCCLS). Protection of laboratory
workers from infectious diseases transmitted by blood and tissue; proposed guideline.
Villanova, PA 1991;7 (9). Order code M29-P.
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Source: http://www.menarinidiag.co.uk/content/download/7999/41628/file/MP-323%20CD24.pdf