Immunohistochemistry on Paraffin Sctions: 10-20 μg/mL (1/20-1/0). Mcrowavepretreatment for antigen retrieval recommended. Suggested positive control: Human spleen. Does not react on routinely processed Frozen Sections. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
实验流程
Immunohistochemitry With Unlabeled Monoclonal Antibodies On Paraffin-EmbeddedTissue Sections1) 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 diluted 1: 10 in Tris buffered saline pH 7. 5, containing0. 1% bovine serum albumin (BSA). Cover sections with NSS and incubate for 15min in ahumid chamber at room temperature (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) 1: 400 diluted inTBS/0. 1%BSA, pH 7. 5 for 30min in a moist chamber, room temperature. Preincubate nowStreptABComplex /HRPO in TBS/0. 1%BSA, pH 7. 5! 10) Rinse sections in TBS, pH 7. 5 for 5min, 2x11) Incubate with StreptABComplex /HRPO for 30min in a humid chamber at roomtemperature12) 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
限制
仅限研究用
溶解方式
Restore by adding 0.5 mL distilled water.
浓度
0.4 mg/mL
缓冲液
PBS, pH 7.2, containing 5 mg/mL BSA as stabilizer and 0.01 % Kathon as preservative.
储存液
Kathon CG
注意事项
This product contains Kathon CG a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
注意事项
Avoid repeated freezing and thawing.
储存条件
4 °C/-20 °C
储存方法
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Allograft Inflammatory Factor-1 is a Ca2+-binding peptide produced by activated macrophages and microglial cells. It has been suggested that AIF-1 expression is associated with chronic inflammatory processes. AIF-1, like the Ca2+-binding peptides MRP8 and MRP14, is expressed by activated macrophages and might participate in a variety of pathogenic processes in the mammalian brain and in chronic transplant rejection. It has been shown to be expressed early and persistently in chronically rejecting cardiac allografts but not in cardiac syngrafts and host hearts. The recently described apoptosis-inducing factor AIF is not related to AIF-1.Synonyms: AIF-1, Allograft inflammatory factor 1, IBA1, Ionized Calcium-Binding Adapter Molecule 1, Protein G1