Immunohistochemistry on Frozen Sections: 10 μg/mL (1/100). Immunohistochemistry on Paraffin Sections: 10 μg/mL (1/100). Microwave pretreatment incitrate buffer is recommended for antigen retrieval in paraffin embedded tissues. Recommended Positive Control: Human tonsil. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protokoll
Immunohistochemistry on paraffin sections: 1) Deparaffinize slides in chloroform for 30min and rehydrate to distilled water2) Antigen retrieval: microwave treatment (700W) for 15min in 300ml 0. 01M citrate bufferpH 6. 0. 3) Incubate slides for 15min in 200ml methanol/4. 8ml 30%H2O24) Rinse slides 2x in distilled water and for 5min in TBS, pH 7. 55) Block with normal swine serum (NSS, Jackson Immunoresearch #014-000-121) diluted1: 10 in Tris buffered saline pH 7. 5, containing 0. 1% bovine serum albumin (BSA,Calbiochem). Cover sections with NSS and incubate for 15min in a humid chamber at roomtemperature (RT). 6) Remove serum from sections (do not rinse)7) Incubate with primary antibody diluted in TBS/0. 1%BSA, pH 7. 5 for 60min in a humidchamber at room temperature8) Rinse sections in TBS, pH 7. 5 for 5min, 2x9) Incubate with secondary antibody (rabbit anti mouse IgG+IgM-biotin, Dako E 0464)1: 400 diluted in TBS/0. 1%BSA, pH 7. 5 for 30min in a moist chamber, room temperature. Preincubate now StreptABComplex /HRPO in TBS/0. 1%BSA, pH 7. 5! 10) Rinse sections in TBS, pH 7. 5 for 5min, 2x11) Incubate with StreptABComplex /HRPO (DAKO, K 0377) for 30min in a humid chamber atroom temperature12) Rinse sections in TBS, pH 7. 5 for 5min, 2x13) DAB-staining Prepare 3,3 - diaminobenzidintetrahydrochloride (DAB) according themanufacturers instructions (e. g. DAB tablets) and incubate slides for 1-15 min. Check colorreaction to avoid overstaining. 14) Rinse sections in TBS, pH 7. 5 for 5min, 2x15) Counterstain in haematoxylin for 1min and rinse in cold tap water for 10min16) Mount slides with coverslips
Beschränkungen
Nur für Forschungszwecke einsetzbar
Rekonstitution
Restore with 0.5 mL distilled water.
Konzentration
0.4 mg/mL
Buffer
Stock solution contains PBS, pH 7.2 with 5 mg/mL BSA as a stabilizer and 0.09 % Sodium Azide as preservative
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. Should this product contain a precipitate we recommend microcentrifugation before use.
Lagerung
4 °C
Informationen zur Lagerung
Store the antibody undiluted at 4-8 °C for one month or at -20 °C for longer.
KNO antikoerper, KNO1 antikoerper, KS antikoerper, cb371 antikoerper, cb529 antikoerper, fc96d06 antikoerper, sb:cb529 antikoerper, wu:fb47h10 antikoerper, wu:fb77g12 antikoerper, wu:fc50f04 antikoerper, wu:fc96d06 antikoerper, wu:fk02h12 antikoerper, kno antikoerper, kno1 antikoerper, collagen type XVIII alpha 1 chain antikoerper, collagen type XVIII alpha 1 chain a antikoerper, collagen, type XVIII, alpha 1 antikoerper, collagen type XVIII alpha 1 antikoerper, COL18A1 antikoerper, col18a1a antikoerper, Col18a1 antikoerper, col18a1 antikoerper
Hintergrund
Endostatin is a powerful inhibitor of endothelial cell proliferation and angiogenesis and shows widespread distribution, mainly in vessel walls and some other basement membrane zones.Synonyms: COL18A1, Endostatin, KNO1