Journal: Scientific Reports
Article Title: Expression and localisation of Rab44 in immune-related cells change during cell differentiation and stimulation
Figure Lengend Snippet: Immunofluorescence analysis of Rab44 and markers for haematopoietic stem cells in the bone marrow. ( a ) The fixed sections of femur containing bone marrow were blocked in PBS containing 5% normal donkey serum. The samples were incubated with rabbit polyclonal anti-Rab44 IgG (1:1,000) as the first antibody followed by fluorescent labelling with Alexa fluor 488-conjugated anti-rabbit IgG and then visualised by confocal laser microscopy. Bar: 20 μm. ( b , c ) The mouse bone was fixed, permeabilised with 0.1% Triton X-100 in PBS, and then allowed to react with antibodies against Rab44 and ( b ) CD117 or ( c ) Sca-1. After washing, the samples were incubated with a fluorescence-labelled secondary antibody and then visualised by confocal laser microscopy. Bar: 5 μm.
Article Snippet: The blots were blocked with 5% nonfat milk solution containing Tris-buffered saline (TBS)/0.1% Tween 20 for 1 h at 25 °C, probed with various antibodies overnight at 4 °C, washed, incubated with horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology, Danvers, MA, USA), and finally detected with ECL-Prime (GE Healthcare Life Sciences, Tokyo, Japan).
Techniques: Immunofluorescence, Incubation, Microscopy, Fluorescence