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.
Protocol
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
Restrictions
For Research Use only
Reconstitution
Restore with 0.5 mL distilled water.
Concentration
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
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing. Should this product contain a precipitate we recommend microcentrifugation before use.
Storage
4 °C
Storage Comment
Store the antibody undiluted at 4-8 °C for one month or at -20 °C for longer.
KNO antibody, KNO1 antibody, KS antibody, cb371 antibody, cb529 antibody, fc96d06 antibody, sb:cb529 antibody, wu:fb47h10 antibody, wu:fb77g12 antibody, wu:fc50f04 antibody, wu:fc96d06 antibody, wu:fk02h12 antibody, kno antibody, kno1 antibody, collagen type XVIII alpha 1 chain antibody, collagen type XVIII alpha 1 chain a antibody, collagen, type XVIII, alpha 1 antibody, collagen type XVIII alpha 1 antibody, COL18A1 antibody, col18a1a antibody, Col18a1 antibody, col18a1 antibody
Background
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