Immunofluorescence analysis of RAB3D was performed using 70% confluent log phase 3T3-L1 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 RAB3D Rabbit Polyclonal Antibody (PA1773) at 2µg/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) 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 represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Mouse, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) A S A S E P P A S P R D A A D Q N(18) of rat RAB3D.|
|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)||2 µg/ml|
|Western Blot (WB)||2.0 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.
|Immunohistochemistry (IHC)||See 2 publications below|
PA1-773 detects recombinant rat and mouse rab 3D. This antibody does not detect rab 3A, 3B, or 3C.
PA1-773 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~29 kDa protein representing recombinant mouse rab 3D.
PA1-773 immunogen is a synthetic peptide corresponding to residues M(1) A S A S E P P A S P R D A A D Q N(18) of rat RAB3D. This sequence is completely conserved in mouse. This immunizing peptide (Cat. # PEP-144) is available for use in neutralization and control experiments.
Rab proteins are low-molecular-weight GTP-binding proteins that form the largest branch of the Ras superfamily of GTPases. Located on the cytoplasmic face of organelles and vesicles, rab proteins are involved in intracellular membrane fusion reactions. Three membrane proteins, synaptosomal associated protein of 25 kDa (SNAP-25), synaptobrevin, and syntaxin, form the core of a ubiquitous membrane fusion machine that interacts with the soluble proteins N-ethylmaleimide-sensitive factor (NSF) and a-SNAP. Rab proteins, in coordination with the core fusion machinery and Munc-18, help to mediate vesicle docking and fusion. There exist over 40 rab proteins that have been described in mammals. Rab 3D has been shown to be present, in part, in the tongue, pancreas, and liver.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Immunohistochemical localization and mRNA detection of Rab3D and/or Rab3B in rat von Ebner's glands, parotid gland, pancreas, and liver.
PA1-773 was used in immunohistochemistry to investigate the distribution of Rab3D/B in rat organs
|Field RB,Kruse DH,Redman RS||The Histochemical journal (33:71)||2001|
Role of actin in regulated exocytosis and compensatory membrane retrieval: insights from an old acquaintance.
PA1-773 was used in immunohistochemistry to investigate the importance of actin in exocytosis and reciprocal membrane retrieval in pancreatic acinar cells
|Valentijn K,Valentijn JA,Jamieson JD||Biochemical and biophysical research communications (266:652)||1999|