Immunofluorescence analysis of Resistin 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 Resistin Rabbit Polyclonal Antibody (PA11050) 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 cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues S(21) S M P L C P I D E A I D K K I K Q D C(39) of mouse Resistin.|
|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)||2 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA1-1050 detects recombinant mouse resistin.
PA1-1050 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~14 kDa protein representing recombinant mouse resistin.
The PA1-1050 immunogen is a synthetic peptide corresponding to residues S(21) S M P L C P I D E A I D K K I K Q D C(39) of mouse Resistin. This peptide (Cat. # PEP-165) is available for use in neutralization and control experiments.
Adipose tissue is an energy reserve in animals and, in nondomestic species, is strictly regulated. Adipose cells produce and secrete numerous physiologically important proteins, such as lipoprotein lipase (LPL), leptin, adipocyte complement related protein of 30 kDa (Acrp30), perilipin, and resistin. Resistin, also known as Adipocyte Secreted Factor (ADSF) and FIZZ3, is a cytokine that is specifically secreted by adipocytes. While the function of resistin is not well understood, it appears to have key paracrine roles and has a suggested role in type 2 diabetes. Studies show that in obese, diabetic mice the levels of serum resistin are increased, which has an antagonistic effect on insulin. Additional studies have shown that resistin causes insulin resistance and glucose intolerance in mice.
Resistin gene expression requires C/EBP alpha, which is necessary and sufficient for expression. Studies show that the coactivators CBP and p300, which have been shown to display histone acetyltransferase (HAT) activity, abundantly acetylate the histones at the resistin promoter suggesting their role in activating chromatin for resistin transcription. Numerous PPAR gamma and RXR ligands have been shown to downregulate resistin expression in adipocytes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
The hormone resistin links obesity to diabetes.
PA1-1050 was used in blocking/activating experiment, immunohistochemistry and western blot to study the role of resistin in linking obesity to diabetes
|Steppan CM,Bailey ST,Bhat S,Brown EJ,Banerjee RR,Wright CM,Patel HR,Ahima RS,Lazar MA||Nature (409:307)||2001|