Immunofluorescence analysis of VAMP4 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with VAMP4 Rabbit Polyclonal Antibody (PA1768) 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 (A27034) 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 represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant rat VAMP-4 protein.|
|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 µg/ml|
|Immunofluorescence (IF)||10 ug/ml|
|Western Blot (WB)||0.5-3 µg/ml|
* 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.
PA1-768 detects VAMP-4 from human and mouse samples.
PA1-768 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~18 kDa protein representing VAMP-4 from PC3 cell extract. Immunofluorescent staining of VAMP-4 in CV-1 cells using PA1-768 results in a staining pattern consistent with VAMP-4 staining.
Immunofluorescence staining with this antibody requires saponin permeabilization.
The PA1-768 antigen is recombinant rat VAMP-4.
The vesicle associated membrane proteins (VAMP) or synaptobrevins are calcium binding proteins specific to eukaryotes. VAMPs, along with synaptosomal associated protein of 25 kDa (SNAP-25) and syntaxin, form the core complex of soluble NSF attachment protein receptor (SNARE) proteins that interact with the soluble proteins N-ethylmaleimide-sensitive factor (NSF) and alpha-SNAP. These membrane associated proteins play a key role in the regulation of vesicle membrane fusion with the plasma membrane. The Clostridium tetani neurotoxin is a metalloprotease with specificity for VAMP. In Alzheimer and quote;s disease, VAMP levels of all isoforms appear to be significantly lowered.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Vesicular transport system in myotubes: ultrastructural study and signposting with vesicle-associated membrane proteins.
PA1-768 was used in immunocytochemistry and western blot to study the morphology of the myotube vesicular transport system
|Tajika Y,Takahashi M,Khairani AF,Ueno H,Murakami T,Yorifuji H||Histochemistry and cell biology (141:441)||2014|
VAMP2 marks quiescent satellite cells and myotubes, but not activated myoblasts.
PA1-768 was used in immunocytochemistry to investigate VAMP2 expression and its cellular distribution during muscle differentiation
|Tajika Y,Takahashi M,Hino M,Murakami T,Yorifuji H||Acta histochemica et cytochemica (43:107)||2010|
An intracellular role for ABCG1-mediated cholesterol transport in the regulated secretory pathway of mouse pancreatic beta cells.
PA1-768 was used in western blot to investigate the role of ABCG1 in subcellular cholesterol distribution in beta cells
|Sturek JM,Castle JD,Trace AP,Page LC,Castle AM,Evans-Molina C,Parks JS,Mirmira RG,Hedrick CC||The Journal of clinical investigation (120:2575)||2010|