MRC-5 cells (fetal lung fibroblasts) were grown on glass coverslips in 6 well dishes and infected for 48 h with VZV. Cells were fixed in 2% Paraformaldehyde and permeabilized with 0.02% Triton X-100 for 1 h at room temperature. Blocked cells for 2 h at RT in 5% non-fat milk plus 2.5% normal goat serum in PBS. Anti-VZV gE mouse monoclonal antibody (red) and anti-Rab5 rabbit polyclonal antibody (green, #PA5029022, 1:100 in PBS) were incubated 2 h at RT (with rocking), then overnight at 4oC (with gentle rocking). After washing with PBS, cells were incubated with goat anti rabbit AlexaFluor488 (1:1250), goat anti mouse AlexaFluor546 (1:1250) and Hoechst 33342 (blue, 1:1000) for 2 h at RT (rocking, in the dark). Washed the cells with PBS. Mounted coverslips to glass and viewed on Zeiss 710 Laser Scanning Confocal Microscope. Images shown are at 400X (upper) or 630X (bottom) total magnification, respectively. Data courtesy of Dr. Grose and quote;s lab.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to a region within amino acids 132 and 215 of Human RAB5A|
|Purification||Antigen affinity chromatography|
|Storage buffer||0.1M tris glycine, pH 7, with 10% glycerol|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:1000|
|Western Blot (WB)||1:500-1:3000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA5-29022 targets RAB5A in WB applications and shows reactivity with Human and mouse samples. PA5-029022 detects Rab5 in IF application with human samples.
The PA5-29022 immunogen is synthetic peptide corresponding to a region within amino acids 132 and 215 of Human RAB5A.
Required for the fusion of plasma membranes and early endosomes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.