Immunohistochemistry on: Frozen sections: 0.5 μg/mL (1/200). Paraffin sections: 4 μg/mL (1/25), Proteinase K pretreatment for antigen retrievalrecommended. Suggested positive control: Human uterus. Antigen Distribution: In contrast to MCA producing tumors, the b-12 related antigen is onlylocated at the MCA producing sites such as glandular cell surfaces or glandular tubuli. InMCA producing tumors, where cells become disorganized, the b-12 antigen is secreted intostromal tissue and blood vessels. (See Table 1. ) Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protokoll
Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature1. Wash in PBS2. Block endogenous peroxidase3. Wash in PBS4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber6. Wash in PBS7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS11. Counterstain with Mayer's hemalum. Protocol with formalin-fixed, paraffin-embedded sections: The whole procedure is performed at room temperature1. Deparaffinize and rehydrate tissue section2. Incubate the tissue section with proteinase K for 7 min. 3. Wash in distilled water4. Block endogenous peroxidase5. Wash in PBS6. Block with 10% normal goat serum in PBS for 30min. in a humid chamber7. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber8. Wash in PBS9. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber10. Wash in PBS11. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 12. Wash in PBS13. Counterstain with Mayer's hemalum.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Rekonstitution
Restore with 0.5 mL distilled water.
Konzentration
0.1 mg/mL
Buffer
PBS, buffer pH 7.2 with 0.05 % Sodium Azide as preservative and 2 mg/mL BSA as stabilizer
Konservierungsmittel
Sodium azide
Vorsichtsmaßnahmen
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung
Avoid repeated freezing and thawing.
Lagerung
4 °C/-20 °C
Informationen zur Lagerung
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.