[Intracellular Polyamine Imaging Reagent] PolyamineRED [FDV-0020]_Funakoshi - 코아사이언스

 

The world's first Intracellular Polyamine Imaging Reagent

PolyamineRED 

Intracellular Polyamine Imaging Reagent

Cat.# FDV-0020 / Size: 0.5mg

Datasheet 

 

본 시약은 세포 내 폴리아민을 특이적으로 이미징하는 시약으로 세포 용해 및 어떠한 전처리 과정이 필요하지 않습니다. 

살아있는 세포에 사용할 수 있기 때문에 세포 내 폴리아민의 검출, 분포 확인 또는 반정량 분석에 적합합니다.

 

PolyamineRED is a reagent for imaging intracellular polyamines specifically without any pretreatment or cell lysis. 

This can be used for live cells, therefore it is suitable for detection or semi-quantitative analysis of intracellular polyamines.

This product has been commercialized with the support of Biofunctional Synthetic Chemistry Laboratory, RIKEN.

 

Background

The polyamine species, such as putrescine, spermidine and spermine etc. and its acetyl derivatives, are some of the essential class of metabolites which have liner alkyl structure with two or more amines.
Polyamines have polycationic properties and show various biological functions.
For example, polyamines interact with DNA and RNA in the nuclear and regulate gene expression. 
Polyamines also interact with negatively charged proteins and control its function. 
The major source of polyamines is amino acid ornithine. In case of mammalian, ornithine is converted to putrescine by ornithine decarboxylase (ODC), followed by synthesizing spermindine and spermine. 
As ODC is highly expressed in cancer cells, polyamines are considered as cancer marker.

Several detection methods of polyamines are developed so far, but, most of them are low-throughput systems using HPLC with polyamine standard compounds. Therefore, biological functions of polyamines have been unclear.

PolyamineRED is the world's first reagent for detecting intracellular polyamines without any pre-treatment and cell lysis. 
This TAMRA (tetramethylrhodamine)-conjugated derivative of glycine propagyl ester specifically reacts with linear primary alkylamine and has cell-penetrating properties, specifically reacts with polyamines inside the cells and labelled polyamines with red fluorescent dye TAMRA.

Features 

• Specifically labels polyamines
• Can be detected by filter set for Rhodamine (Ex./Em. = 560 / 585 nm)
• Cell permeable : compatible with live cell imaging
• No pretreatments required
• Can do semi-quantification of total polyamines amount (*) by intracellular fluorescence intensity.

* Only total polyamines, not for each polyamine.

 

Chemical Information

• Molecular weight : 611 g/mol
• Solubility : Soluble in DMSO
• Fluorophore : TAMRA (red fluorescent dye)

Example Data 

Fig. 1 Polyamine imaging in cancer or non-cancer cells
Three cancer cell lines (MCF7, MDA-MB-231 and SK-BR-3) and two non-cancer cells (MCF10A and human lymphocyte) were treated with 30 μM of PolyamineRED for 10 min. After incubation, cells were washed three times by PBS, followed by DAPI staining and formalin fixation. Images were obtained at Ex/Em=560 nm/585 nm for TAMRA and at Ex/Em=358 nm/461 nm for DAPI. TAMRA fluorescence was detected in cancer cell lines.

Fig.2 Intracellular distribution of polyamines in MDA-MB-231 cancer cell lines
MDA-MB-231 cells were treated with 30 μM of PolyamineRED for 10 min. After incubation, cells were washed three times by PBS, followed by DAPI-staining and formalin fixation. Images were obtained at Ex/Em=560 nm/585 nm for TAMRA and at Ex/Em=358 nm/461 nm for DAPI. Major TAMRA signal was detected in nucleus. This indicates polyamines in MDA-MB-231 are mainly localized in nucleus.

 

Amine	Reaction
Spermine	82%
Spermidine	78%
Putrescine	66%
Epinephrine	<1%
Lysine	2%

Table Selectivity of glycine propagyl ester to polyamines
Benzyloxycarbonyl glycine propagyl ester as a model molecule was selectively reacted with polyamines. 
Reactant of epinephrine, an example of monoamine, and lysine, an example of amino acid, were rarely detected. 
Reactivity for polyamines depends on the length of polyamine and double linkage products were observed from spermine (4 amino groups) and spermidine (3 amino groups). 
Data is cited from K. Vong, K. Tsubokura, Y. Nakao, T. Tanei, S. Noguchi, S. Kitazume, N. Taniguchi, K. Tanaka, Chem. Commun., 53, 8403-8406 (2017).

 

Reference 

K. Vong, K. Tsubokura, Y. Nakao, T. Tanei, S. Noguchi, S. Kitazume, N. Taniguchi, K. Tanaka, Chem. Commun., 53, 8403-8406 (2017).

 

NOTICE : ALL PRODUCTS ON THIS E-MAIL ARE FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC USE.

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