The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
Use a concentration of 10 µg/ml.
Use at an assay dependent concentration. PubMed: 28754163
Chitinase-3-like protein 3 (Ym1) is a lectin that binds saccharides with a free amino group, such as glucosamine or galactosamine; binding to oligomeric saccharides is much stronger. It also binds chitin and heparin. This protein has been implicated in inflammation and allergy.
Immunocytochemistry/ Immunofluorescence - Anti-Chitinase 3 like protein 3 antibody (ab93034)
ICC/IF image of ab93034 stained RAW 246.7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab93034 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Chitinase 3 like protein 3 antibody (ab93034)
All lanes : Anti-Chitinase 3 like protein 3 antibody (ab93034) at 1 µg/ml
Lane 1 : Mouse Bone Marrow Tissue Lysate Lane 2 : Spleen (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 44 kDa Observed band size : 44 kDa Additional bands at : 26 kDa,37 kDa. We are unsure as to the identity of these extra bands.
Ge R et al. Choroid plexus-cerebrospinal fluid route for monocyte-derived macrophages after stroke. J Neuroinflammation14:153 (2017).
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Katsuyama E et al. Interleukin-1 receptor-associated kinase-4 (IRAK4) promotes inflammatory osteolysis by activating osteoclasts and inhibiting formation of foreign body giant cells. J Biol Chem290:716-26 (2015).
Read more (PubMed: 25404736) »