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Vacuole membrane protein 1 polyclonal antibody

 
ALX-215-068-C200 200 µg 301.00 USD
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VMP1 is a stress-induced protein that, when overexpressed, promotes formation of intracellular vacuoles followed by cell death. May be involved in the cytoplasmic vacuolization of acinar cells during the early stage of acute pancreatitis.VMP1 is a transmembrane protein that co-localizes with LC3, a marker of the autophagosomes. It interacts with beclin-1, a mammalian autophagy initiator, through the VMP1-Atg domain, which is essential for autophagosome formation. VMP1 endogenous expression co-localizes with LC3 in pancreas tissue undergoing pancreatitis-induced autophagy.

Product Specification

Alternative Name:VMP1, TMEM49
 
Host:Rabbit
 
Immunogen:Synthetic peptide corresponding to aa 386-406 of human vacuole membrane protein 1 (VMP1).
 
UniProt ID:Q96GC9
 
Species reactivity:Human, Mouse, Rat
 
Crossreactivity:Ovalbumin
 
Applications:ICC, IP, WB
 
Recommended Dilutions/Conditions:Immunocytochemistry (1:200)
Immunoprecipitation (5µg/mg lysate)
Western Blot (1:100-1:1,000)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
 
Formulation:Liquid. Purified serum containing 0.2% sodium azide.
 
Handling:Avoid freeze/thaw cycles.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
Scientific Background:VMP1 is a transmembrane protein that co-localizes with LC3, a marker of the autophagosomes. It interacts with beclin-1, a mammalian autophagy initiator, through the VMP1-Atg domain, which is essential for autophagosome formation. VMP1 endogenous expression co-localizes with LC3 in pancreas tissue undergoing pancreatitis-induced autophagy.
 
Protocol:Standard Procedure for Immunocytochemistry
  1. Grow cells on glass coverslips.
  2. Rinse briefly in phosphate-buffered saline (PBS).
  3. Fix cells with 4% paraformaldehyde in PBS, pH 7.4 for 10 min at room temperature.
  4. Wash cells three times with PBS.
  5. Incubate the samples for 5 min with PBS containing 0.1% Triton X-100.
  6. Wash 3 x 5 min with PBS.
  7. Incubate cells with 1% BSA in PBS for 1 hour at room temperature (this time can be adjusted to the study system).
  8. Incubate with primary antibody diluted in PBS containing 1% BSA in a humidified chamber for 1hr at room temperature. VMP1 antibody dilution 1:200.
  9. Wash 1 x 5 min with 0.05% Tween-20 in PBS.
  10. Wash 2 x 5 min with PBS.
  11. Incubate with secondary antibody in PBS containing 1% BSA for 1 h at room temperature in the dark. Alexa Fluor conjugated antibodies dilution 1:1000.
  12. Wash 2 x 5 min with 0.05% Tween-20 in PBS.
  13. Wash 3 x 5 min with PBS.
  14. (OPTIONAL) Incubate cells with DAPI (DNA stain) for 1 min.
  15. Rinse with PBS.
  16. Mount coverslip with a drop of mounting medium.
  17. Seal coverslip with nail polish to prevent drying and analyze under a microscope.
  18. Store coverslips in the dark at -20°C.
 

Product Literature References

Autophagy and VMP1 expression are early cellular events in experimental diabetes.: D. Grasso, et al.; Pancreatology 9, 81 (2009), Abstract;
A novel mammalian trans-membrane protein reveals an alternative initiation pathway for autophagy: M. Vaccaro, et al.; Autophagy 4, 1 (2008), Abstract;
The pancreatitis-induced vacuole membrane protein 1 triggers autophagy in mammalian cells: A. Ropolo, et al.; J. Biol. Chem. 282, 37124 (2007), Abstract;

General Literature References

Autophagy dysfunction and ubiquitin-positive protein aggregates in Dictyostelium cells lacking Vmp1: J. Calco-Garrido & R. Escalante; Autophagy 6, 99 (2010), Abstract;
The TP53INP2 Protein Is Required for Autophagy in Mammalian Cells: J. Nowak, et al.; Mol. Biol. Cell 20, 870 (2009), Abstract;
TP53INP2 is the new guest at the table of self-eating: J. Novak & J.L. Iovanna; Autophagy 5, 383 (2009), Abstract;

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