Asante Calcium Red Info Packet

Download the updated version (May 28 2010) of the ACR Info Packet

 

 

Info Packet Contents:

Asante Calcium Red - Next Generation Calcium Indicator
One indicator, two excitation-dependent emission response modes 

emission ratiometric mode (Figure 1)

non-ratiometric mode (Figures 2 and 3)
Asante Calcium Red Properties
 Kd of ~300 to 400 nM
 pKa of ~5.5
 More than twice as bright as Fura-Red; not as bright as Fluo dyes, however
 AM ester is nonfluorescent
 Potassium Salt product specifications
 AM ester product specifications
 Potassium Salt MSDS
 AM ester MSDS
 AM ester loading in rat cardiac myocytes and rat vagal sensory neurons

Loading ACR

From a customer conversation with our collaborator regarding the loading of Asante calcium Red:

"...In the case of T cells, the AM ester form must be used (T cells are lymphocytes, which are quite small). In the case of larger cells, one can sometimes inject the salt form into individual cells through a micro-needle (tip opening less than 1 micrometer diameter). Microinjection is laborious and inefficient and is not generally the loading method of choice..."

ACR-AM loading procedures:

In rat cardiac myocytes1:
Acutely dissociated rat cardiac myocytes were incubated for 45 min at room temperature with Tyrode’s solution containing 10 uM Asante Calcium Red (AM) and 0.02% (wt/vol) Pluronic F-127. The cells were then transferred to fresh Tyrode’s solution and allowed to stand for an additional 45 min to ensure complete hydrolysis of the AM ester.

In rat vagal sensory neurons2:
The inferior vagal (nodose) ganglia from a Sprague-Dawley rat were dissociated enzymatically. The yield of nodose neurons was suspended in Leibovitz L-15 medium supplemented with 10% (v/v) fetal bovine serum and penicillin-streptomycin and plated onto No. 1 glass coverslips. Neurons were incubated for 50 min at room temperature with 3 uM Asante Calcium Red (AM) ester. Thereafter, the cells were washed and maintained in fresh L-15 medium for 40 min to permit intracellular enzymatic hydrolysis of the AM ester to proceed to completion.

 

 

AM ester loading conditions and confocal imaging of sensing of calcium transients by Asante Calcium Red were contributed by:

1 Rat cardiac myocytes:
BM Hagen, WJ Lederer, JPY Kao
Center for Biomedical Engineering & Technology
University of Maryland, Baltimore
Medical Biotechnology Center
University of Maryland Biotechnology Institute
 

2 Rat vagal sensory neurons:
G. Li and J.P.Y. Kao
Center for Biomedical Engineering & Technology
University of Maryland, Baltimore
Medical Biotechnology Center
University of Maryland Biotechnology Institute