Immunofluorescence analysis of p115 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with p115 Rabbit Polyclonal Antibody (PA1006) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Mouse, Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) N F L R G V M G G Q S A G P Q H(17) of rat p115.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2-3 µg/mL|
|Immunofluorescence (IF)||2-3 µg/mL|
|Western Blot (WB)||1:500|
* 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.
PA1-006 detects p115 in rat and mouse samples.
PA1-006 has been successfully used in Western blot procedures. By Western blot, this antibody detects a ~115 kDa protein representing p115 in rat and mouse liver samples.
The PA1-006 immunogen is a synthetic peptide corresponding to residues M(1) N F L R G V M G G Q S A G P Q H(17) of rat p115. This sequence is conserved across many species including mouse and human. PA1-006 immunizing peptide (Cat. # PEP-276) is available for use in neutralization and control experiments.
p115 (Vesicle docking protein p115) is a peripheral membrane protein that is located on the Golgi complex. p115 exists as a homodimer with two globular heads, an extended coiled-coil tail, followed by an acidic domain at the extreme C terminus. p115 is homologous to a yeast protein, Uso1p, which is required for ER to Golgi transport. p115 likely plays an important role in vesicle transportation from the ER to the cis-Golgi comparments.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.