AbD Serotec
Hamster anti-mouse CD11c
MCA1369GA
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Our research looks at immune cells such as macrophages, lymphocytes, and dendritic cells. One of our experiments required us to identify CD11c+ cells in the mouse spleen (typically dendritic cells and to some extent, even macrophages double-stain with F4/80 surface marker) using this antibody and a secondary fluorescent antibody.
Immunofluorescence
8-10 weeks Mouse spleen cryosections - control animals (no experimental treatment)
Tried 1:10 - 1:100 dilution, at 4 degrees C in the dark overnight
Blocking buffer used was PBS with 0.05% Tween 20, 1% BSA, 5% Host serum and 0.3 M Glycine
Anti-hamster IgG FITC (Catalog No. ab5739 from abcam), tried 1/20-200 for 1 hour at room temperature in the dark
Used Hoechst 33343 for nuclear staining
FITC filter in any standard epifluorescence microscope
As seen in the image, the overlay indicates no CD11c staining after subtracting using the IgG control staining around cells (made visible via the nuclear stain - Hoechst 33342). Since multiple dilutions were attempted, and the fact that the sample is a positive sample for CD11c detection, it was surprising to see no detection at all. Also, the secondary antibody has been used before for other applications.
None
Did not work for detection in mouse spleen.
This particular product did not work for our experiments to detect CD11c+ cells. Other options or methods should be considered.