Immunofluorescence analysis of Perilipin A was performed using 70% confluent 3T3-L1 cells differentiated with StemPro Adipogenesis Supplement (Product # A10065-01) for 5 days. 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 Perilipin Rabbit Polyclonal Antibody (PA1-1051) 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) 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 cytosolic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Mouse, Rat|
|Published species reactivity||Rat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the residues C E(502) P I L G R T Q Y S Q L R K K S(517) of rat Perilipin A.|
|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)||2 µg/ml|
|Western Blot (WB)||1:1000-1:4000|
* 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.
PA1-1051 detects perilipin A from rat and 3T3-L1 cell extract.
PA1-1051 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~62 kDa protein representing perilipin A from 3T3-L1 cell extract
The PA1-1051 immunogen is a synthetic peptide corresponding to the residues C E(502) P I L G R T Q Y S Q L R K K S(517) of rat Perilipin A. This sequence is 93% conserved in humans. The PA1-1051 immunizing peptide (Cat. # PEP-169) is available for use in neutralization and control experiments.
Adipose tissue is an energy reserve in animals and is strictly regulated in nondomestic species. Adipose cells produce and secrete numerous physiologically important proteins, such as lipoprotein lipase (LPL), leptin, adipocyte complement related protein of 30 kDa (Acrp30), resistin, and perilipin. Perilipin is an intracellular neutral lipid droplet protein that is hormonally regulated. This protein is localized exclusively to the surface of lipid droplets. In response to lypotic stimuli, perilipin is phosphorylated by protein kinase A. Once activated, perilipin has inhibitory affects upon hormone-sensitive lipase (HSL), a protein that mediates the hydrolysis of triacylglycerol, the major form of stored energy in the body.
Perilipin expression is limited to adipocytes and steroidogenic cells. There are currently two known isoforms, Perilipin A and B. Both of these proteins are encoded by a single-copy gene and are the result of differential splicing events.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Suppression of adipose lipolysis by long-chain fatty acid analogs.
PA1-1051 was used in western blot to study the mechanism by which long-chain fatty acid analogs suppress adipose lipolysis
|Kalderon B,Azazmeh N,Azulay N,Vissler N,Valitsky M,Bar-Tana J||Journal of lipid research (53:868)||2012|
DGAT enzymes are required for triacylglycerol synthesis and lipid droplets in adipocytes.
PA1-1051 was used in western blot to study the role of DGAT enzymes in triacylglycerol synthesis and lipid droplets in adipocytes
|Harris CA,Haas JT,Streeper RS,Stone SJ,Kumari M,Yang K,Han X,Brownell N,Gross RW,Zechner R,Farese RV||Journal of lipid research (52:657)||2011|
Medium-chain Fatty acids attenuate agonist-stimulated lipolysis, mimicking the effects of starvation.
PA1-1051 was used in western blot to investigate the role of medium-chain Fatty acids during agonist-stimulated lipolysis
|Lei T,Xie W,Han J,Corkey BE,Hamilton JA,Guo W||Obesity research (12:599)||2004|
|Mouse||Not Cited||Perilipin A increases triacylglycerol storage by decreasing the rate of triacylglycerol hydrolysis.||Brasaemle DL,Rubin B,Harten IA,Gruia-Gray J,Kimmel AR,Londos C||The Journal of biological chemistry (275:38486)||2000|
|Rat||Not Cited||Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes.||Clifford GM,Londos C,Kraemer FB,Vernon RG,Yeaman SJ||The Journal of biological chemistry (275:5011)||2000|
||Perilipin A increases triacylglycerol storage by decreasing the rate of triacylglycerol hydrolysis.||Brasaemle DL,Rubin B,Harten IA,Gruia-Gray J,Kimmel AR,Londos C||The Journal of biological chemistry (275:38486)||2000|