Anti-Rab5 antibody - Drosophila Early Endosome Marker (ab31261)

Immunocytochemistry/ Immunofluorescence - Rab5 antibody - Drosophila Early Endosome Marker (ab31261)

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).

This image is courtesy of an anonymous Abreview

Western blot - Rab5 antibody - Drosophila Early Endosome Marker (ab31261)

Anti-Rab5 antibody - Drosophila Early Endosome Marker (ab31261) at 1 µg/ml + Schneider L2 (Fruit fly (Drosophila melanogaster)) 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 - Rab5 antibody - Drosophila Early Endosome Marker (ab31261)

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.

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