Immunofluorescence analysis of Leptin was performed using 70% confluent log phase 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 Leptin Rabbit Polyclonal Antibody (PA1051) 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the undifferentiated cells that have lower level of Leptin expression. Panel f represents the no primary control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Sheep, Rat|
|Published species reactivity||Avian, Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues Q(25) K V Q D D T K T L I K T I V T R I N D(44) of mouse Leptin.|
|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 *|
|ChIP assay (ChIP)||1.5 µg/10^6 cells|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||1:50 - 1:200|
|Western Blot (WB)||1:200 - 1:1000|
* 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.
|Western Blot (WB)||See 4 publications below|
PA1-051 detects leptin from human, mouse, sheep and rat samples.
PA1-051 has been successfully used in Western blot, immunocytochemistry and immunohistochemistry experiments. By Western blot, this antibody detects an ~16 kDa protein representing leptin from 3T3-L1 cell extract. Immunohistochemical staining of leptin in sheep brain with PA1-051 results in staining of the ventromedial hypothalamus.
The PA1-051 immunogen is a synthetic peptide corresponding to residues Q(25) K V Q D D T K T L I K T I V T R I N D(44) of mouse Leptin. This sequence is completely conserved between mouse and bovine.
Leptin, the obese (ob) gene product, is a small protein expressed in and secreted from adipose tissue of normal rodents. Studies suggest leptin acts as a circulating hormone capable of regulating body-weight homeostasis and energy balance. One possible target tissue for leptin is the hypothalamus, a proposed control center for satiety and energy expenditure.
Ob gene knockout mice are characterized by several metabolic abnormalities including hyperglucocorticoidemia, hyperinsulinemia and insulin resistance, hyperglycemia, altered central nervous system activity, reduced metabolic rate of brown adipose tissue, and a large increase in white adipose tissue. Studies show that the administration of recombinant leptin to ob knockout mice reduces food intake and increases energy expenditure.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Pressure mediated hypertrophy and mechanical stretch up-regulate expression of the long form of leptin receptor (ob-Rb) in rat cardiac myocytes.
PA1-051 was used in western blot to study the upregulation of cardiac expression of the long form of the leptin receptor in response to pressure overload hypertrophy and mechanical stretch
|Matsui H,Yokoyama T,Tanaka C,Sunaga H,Koitabashi N,Takizawa T,Arai M,Kurabayashi M||BMC cell biology (13:null)||2012|
Endoplasmic reticulum stress-induced CHOP activation mediates the down-regulation of leptin in human neuroblastoma SH-SY5Y cells treated with the oxysterol 27-hydroxycholesterol.
PA1-051 was used in western blot to investigate the mechanism of leptin down-regulation in 27-hydroxycholesterol-treated human neuroblastoma cells
|Marwarha G,Dasari B,Ghribi O||Cellular signalling (24:484)||2012|
Leptin reduces pathology and improves memory in a transgenic mouse model of Alzheimer's disease.
PA1-051 was used in western blot to investigate the effect of leptin treatment on histopathology and memory capability in Alzheimer disease model
|Greco SJ,Bryan KJ,Sarkar S,Zhu X,Smith MA,Ashford JW,Johnston JM,Tezapsidis N,Casadesus G||Journal of Alzheimer's disease : JAD (19:1155)||2010|
Leptin is synthesized in the liver and adipose tissue of the dunlin (Calidris alpina).
PA1-051 was used in western blot to investigate the localization of leptin expression in the dunlin
|Kochan Z,Karbowska J,Meissner W||General and comparative endocrinology (148:336)||2006|