Immunofluorescence analysis of Visfatin was done on 70% confluent log phase U87MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Visfatin Rabbit Polyclonal Antibody (PA11045) at 1ug/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 (Product # 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 is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues S (472) F D E I R K N A Q L N I E L E A A H H (491) of human Visfatin.|
|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)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1:250-1:1000|
* 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.
PA1-1045 detects Visfatin (PBEF) in human samples.
PA1-1045 has been successfully used in Western blot procedures. In Western blot analysis of human adipose tissue lysate this antibody detects a ~52 kDa protein representing Visfatin.
The PA1-1045 immunogen is a synthetic peptide corresponding to residues S (472) F D E I R K N A Q L N I E L E A A H H (491) of human Visfatin.
The recently discovered adipocytokine Visfatin is a visceral fat specific protein which has been shown to exert insulin-mimetic effects in insulin-sensitive tissues such as liver, muscle and fat. Visfatin can bind to the insulin receptor and visfatin plasma levels increase with obesity. Based on their properties, adipocytokines are likely to play critical roles in metabolic syndromes such as obesity and diabetes.
IP-MS enrichment of NAMPT (LFQ intensity): NAMPT was enriched 1231-fold from MCF7 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and NAMPT antibody (Part No. PA1-1045). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
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