[AMPA receptor live cell imaging reagent] LiveReceptor AMPAR (AMPA Receptor) [FDV-0018A]_Funakoshi - 코아사이언스

AMPA receptor live cell imaging reagent

LiveReceptor AMPAR

Specific labeling regent for synaptic AMPA type glutamate receptor

Cat.#  FDV-0018A / Size: 10ug

"LiveReceptor AMPAR"은 세계 최초로 출시된 살아있는 신경세포 상의 시냅스 뉴런 AMPA 타입 클루타메이트 수용체 특이적 형광 염색 표지 시약입니다.

  

LiveReceptor AMPAR is the world-first reagent which can specifically label AMPA-type glutamate receptors (AMPARs) with fluorescein dye on living neurons. 
This reagent is based on a unique chemical reaction and can accomplish labeling of AMPARs keeping its ligand-binding pockets and receptor functions. 
LiveReceptor AMPAR is a powerful tool to investigate physiological functions of AMPAR by monitoring receptor trafficking of endogenous AMPARs on live cells. No complicated protocol and genetic manipulation are required.

Generally, fluorescein direct conjugated ligand has been used for labeling receptors, however, such ligands easily detach from ligand binding pocket, and then signal will be faded out. (See below image)

 

Our "LiveReceptor" has a linker of acyl imidazole between fluorescein and ligand. While ligand is binding to receptor, nucleophillic amino acids attacks acyl imidazole and changes its chemical structure. By this method, receptors are directly labeled and keep fluorescence even after the ligand is detached from binding pocket.

This product is commercialized by the research result of Professor Hamachi, at Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University.

Features  

• Can label fluorecein to endogeneous AMPAR
• Highly specific to AMPAR (GluA2-4 subunit). Other ion channel type glutamate receptors will not be labeled.
• Can observe signal for 1 to 4 hours after adding LiveReceptor to medium.
• No cell permiability - only AMPAR on cell surface can be labeled.
• Ion channel function is mainteined after labeling
• Low cytotoxisity under receommended concentration (1 μM)

Chemical Information  

• Formulation : C51H43F3N8O15
• Molecular weight : 1064.92 g/mol
• Visibility : Orange lyophilized powder
• Solubility : Soluble in DMSO *This compound has high water-solubility but it can be easily degraded in water and culture medium. Please avoid storage in water
• Excitation/ Emission: 495/515 nm *Compatible with FITC filter

* PFQX : 6-pyrrolyl-7-trifluoromethyl-quinoxaline-2,3-dione. This is affinity ligand for AMPAR.

Application 

• Live cell imaging
• Immunocytochemistry with specific antibodies
• Immunoprecipitation with anti-fluorescein antibody
• Immunoblotting with anti-fluorescein antibody
• Drug screening for competitive AMPAR antagonist

Example Data

 

Fig.1 Live cell imaging of labelled endogenous
AMPARs in cultured neurons Cultured hippocampal neurons were treated with 1 μM of LiveReceptor AMPAR (in left) or Fluorescein-conjugated ligand as negative control (in right) for 1 hour at 17C and washed out three times with the basal medium. Dendritic spin-like punctual structures were observed on live cells by specifically LiveReceptor. (scal bars, 10 μm)

Fig.2 Live cell imaging of labelled endogenous AMPARs in cultured slice tissue
Acutely prepared hippocampal slices were treated with 1 μM of LiveReceptor AMPAR for 1 hour at 17C in the absence (in left) or presence (in right) of 10 μM NBQX, a potent inhibitor of AMPAR, and washed out three times with the basal medium. Dendritic spin-like punctual structures were observed on live cells and the signal was clearly disappeared by NBQX. (scal bars, 5 μm)

Fig.3 Staining of AMPARs in deep tissue in cultured slice tissue
Cultured slice tissue of mouse was stained by LiveReceptor AMPAR and fixed. After fixation, stained by anti-GluA2/3 antibody. x-y dimension shows a good concordance between LiveReceptor AMPAR and anti-GluA2/3 antibody.
However, x-z dimension shows that anti-GluA2/3 antibody stained the surface of slice tissue only. LiveReceptor AMPAR penetrates into deep tissue and can label them. (Black arrowhead : Top of sliced tissue / White arrowhead : Bottom of sliced tissue)

* FOR RESEARCH USE onLY. NOT FOR DIAGNOSTIC USE.

References 

1. Wakayama, et al., Nat. Commun., 8, 14850 (2017). [PMID : 28387242 ] Chemical labeling for visualizing native AMPA receptors in live neurons.

2. Fujishima, et al., J. Am. Chem. Soc., 134, 3961～3964 (2012).

3  Miki, et al., Chem. Biol., 21, 1013～1022 (2014).

코아사이언스 coresciences innovative cell technologies Funakoshi Inc. Japan 후나코시 한국 대리점 시냅스 뉴런 AMPA 타입 클루타메이트 수용체 특이적 염색 표지 시약