Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody - Drosophila Early Endosome Marker (ab31261)This image is courtesy of an anonymous Abreview
ab31261 staining 3rd instar wing imaginal disc cells of fruit fly (Drosophila melanogaster). Cells were formaldehyde fixed and blocked with 5 % serum for 2 hours at 22°C prior to incubating with the primary antibody, diluted 1/500, for 16 hours at 4°C. A Cy3® conjugated donkey anti-rabbit IgG (H+L) antibody, diluted 1/500, was used as the secondary.
In the image, the nuclei of some cells are marked by GFP (green). The punctate, non-nuclear staining pattern is indicative of endosomes (similar to other endosomal markers such as Hrs).
Western blot - Anti-Rab5 antibody - Drosophila Early Endosome Marker (ab31261)
Anti-Rab5 antibody - Drosophila Early Endosome Marker (ab31261) at 1 µg/ml + Schneider L2 whole cell lysate (ab14893) at 10 µg
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 26 kDa Observed band size: 26 kDa Additional bands at: 135 kDa, 145 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
ab31261 detects a strong 26 kDa band in Drosophila lysate consistent with expected molecular weight of Rab5. In some batches of this antibody, we have observed additional bands at ~135, 145 and 70 kDa, the identity of which are unknown.
Immunocytochemistry/ Immunofluorescence - Anti-Rab5 antibody - Drosophila Early Endosome Marker (ab31261)Image from Huang HR et al, Proc Natl Acad Sci U S A. 2010 May 4;107(18):8322-7. Epub 2010 Apr 19, Fig S2. DOI 10.1073/pnas.1004031107
ab31261 staining Rab5 in Drosophila S2 cells by Immunocytochemistry/ Immunofluorescence. S2 cells were fixed in 4% paraformaldehyde
for 10 minutes and then permeablized by incubating
with 0.2% Triton X-100 in PBS for 5 minutes at room temperature.
After blocking, cells were incubated overnight with ab31261 diluted in PBS containing 10% horse serum and
0.2% Triton X-100. A Fluorescein-conjugated anti-rabbit IgG was used as the secondary.