Anti Iba1, Monoclonal Antibody (NCNP24)

 Anti Iba1 for Immunocytochemistry (Wako Cat.#019-19741) Application for paraffin sections 1) Deparaffinization 100% xylene for 5 min, 3 times 100% ethanol for 5 min, 2 times 90% ethanol for 2 min 70% ethanol for 2 min dH2O, 3 minutes 2) Antigen retrieval 10mM citrate buffer(pH6.0), 90℃, 9min 3) Wash with PBS, 5min, 2 times. 4) Incubate with blocking buffer (2.5% normal goat serum in PBS) , 20min, at room temperature (RT). 5) Wash with PBS, 5min, 2 times. 6) Incubate with anti Iba1 antibody (1:1,000)(Wako Catalog No. 019-19741) in the blocking buffer overnight at 4℃. 7) Wash with PBS, 5min, 3 times. 8) Incubate with Alexa Fluor 488, anti-Rabbit IgG Antibody (1:200) in the blocking buffer for 2hours at RT. 9) Wash with PBS, 5min, 3 times. 10) DAPI staining 11) Mount Application for frozen sections 1) Rat or mouse was perfusion-fixed with 4% paraformaldehyde, replaced sucrose, and prepared 50μm brain section by microtome. 2) Wash with 0.3 % TritonX-100 in PBS, 5 min×3. 3) Incubate with blocking buffer (1 % BSA, 0.3 % TritonX-100/PBS), 2hours, RT. 4) Incubate with Anti Iba1 Antibody (1:1,000) (Wako Catalog No. 019-19741) in the blocking buffer(1 % BSA, and 0.3 % TritonX100 in PBS), over night , 4 ℃. 5) Wash with 0.3 % TritonX-100 in PBS, 5 min×3. 6) Incubate with AlexaFluor488 anti-rabbit IgG(1:200) in the blocking buffer(1 % BSA, and 0.3 % TritonX-100 in PBS), 2hour, RT. 7) Wash with 0.3 % TritonX-100 in PBS, 5 min×3. 8) Mount