Antibodies against Anti Iba1 Rabbit - wako-chem.co.jp

1 Antibodies against Macrophage / Microglia-specific protein iba1 . anti iba1 , Rabbit For Immunocytochemistry Wako Cat. No. 019-19741 (50 g (100 L)). For Western Blotting Wako Cat. No. 016-20001 (50 g (100 L)). Calcium ions are known to be one of the most important signal mediators in all cells including central nervous system (CNS) cells. Calcium ions exert their signaling activity through association with various calcium binding proteins, many of which are classified into a large protein family, the EF hand protein family. iba1 is a 17-kDa EF hand protein that is specifically expressed in macrophages/ microglia and is upregulated during the activation of these cells. Wako supplies Rabbit polyclonal Antibodies were raised against a synthetic peptide corresponding to the iba1 carboxy-terminal sequence, which was conserved among human, rat and mouse iba1 protein sequences. These Antibodies are specifically reactive to microglia/.

2 Macrophages, are appropriate for immuno-double staining of brain tissues and cell culture in combination with monoclonal antibody to GFAP, which specifically reacts to astrocyte. Specificity: Specific to human, mouse and rat microglia and macrophages, but not cross-reactive with neurons and astrocytes. Immuno-double staining of rat primary mixed culture cells and a phase-contrast image of the same filed Green: iba1 , which reacts to anti iba1 antibody (Wako Cat. #019-19741). Red: astrocyte, which reacts to anti GFAP, mAb (Data was provided by Dept. of Neurochemistry, National Institute of Neuroscience (Japan).). 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( ), 90 , 9min 3) Wash with PBS, 5min, 2 times.

3 4) Incubate with blocking buffer ( 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 50 m brain section by microtome. 2) Wash with % TritonX-100 in PBS, 5 min 3. 3) Incubate with blocking buffer (1 % BSA, % 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 % TritonX- 100 in PBS), over night , 4 . 5) Wash with % TritonX-100 in PBS, 5 min 3.

4 6) Incubate with AlexaFluor488 anti - Rabbit IgG(1:200) in the blocking buffer(1 % BSA, and % TritonX-100 in PBS), 2hour, RT. Green: iba1 , which reacts to anti iba1 antibody 7) Wash with % TritonX-100 in PBS, 5 min 3. (Wako Cat. #019-19741). 8) Mount Wako Pure Chemical Ind., Ltd. Wako Chemicals USA, Inc. Wako Chemicals GmbH. 1-2, Doshomachi 3-Chome Toll-Free ( only): 1-877-714-1920 Europe Office Chuo-Ku, Osaka 540-8605, Japan Head Office (Richmond, VA): Fuggerstra e 12, D-41468. Tel: 81-6-6203-3741 Tel: 1-804-714-1920/Fax: 1-804-271-7791 Neuss, Germany Fax: 81-6-6203-1999 Boston Sales Office (Cambridge, MA): Tel: 49-2131-311-0. Online Catalog: Tel: 1-617-354-6772/Fax: 1-617-354-6774. Fax: 49-2131-311100.