Secreted alkaline phosphatase

Tobacco NT-1 suspension cells Human secreted alkaline phosphatase Becerra-Arteaga et al., 2006 27 mg/L Batch culture... 

E. Kelso, J. McLean and M.F. Cardosi, Electrochemical detection of secreted alkaline phosphatase implications to cell based assays, Electroanalysis, 12 (2000) 490-494. 

Fig. 12.14 Left Structures of mono-substituted alkylpolyamines (AKla-5a), two bri-substituted analogues (AK6b, 7b) were also synthesized and examined for SAR confirmation. Right Inhibition of LPS (100 ng/mL)-induced NF-kB induction in HEK-293 cells stably transfected with Tlr4, MD-2, CD 14, and an NF-icB-secreted alkaline phosphatase reporter gene construct. Polymyxin B was used as the positive control...

Although secreted alkaline phosphatase (SEAP) does not allow the monitoring of intracellular location, there are a number of sensitive chemilumi-nescense-based assays for SEAP, allowing very low levels of gene expression to... 

Alkaline phosphatase from E. coli and secreted alkaline phosphatase (SEAP) from mammalian cells are two examples of secreted reporter proteins (Berger, 1988 Manoil et al., 1990 Yang et al., 1997). Although these reporters are used less frequently, they have the evident advantage that no cell extracts are required. Different substrates allow detection with different detection tools. 

Yang, T.T., Sinai, P., Kitts, P.A. and Kain, S.R. (1997). Quantification of gene expression with a secreted alkaline phosphatase reporter system. Biotechniques, 23, 1110-1114. 

Decreasing the membrane fluidity also affects secretion. Pagds et al. (1978) varied fluidity by changing the temperature at which E. coli spheroplasts were grown. They found that the amount of secreted alkaline phosphatase (as a fraction of total synthesized protein) dropped dramatically from 29 to 13°C, while the amounts found in the cytoplasm... 

A very recent publication speaks of yet another class of sequence elements that may mediate and maintain high-level productivity in mammalian cells upon gene transfer Highly conserved anti-repressor elements of 1500-2000 base pairs have been identified and cloned from human genomic libraries and inserted into expression vectors. Some of these elements have shown to allow the generation of CHO cell clones producing secreted alkaline phosphatase (SEAP) at 50-80 pg per cell per day [60]. 

FP - fluorescence polarization FI - fluorescence intensity CNG2 - cAMP gated ion channel 2 CRE - cAMP response element SRE - serum response element SEAP - secreted alkaline phosphatase. For further details see Refs. [111-113]... 

It is possible to build up a picture of mineralization from these various concepts which has not yet been completely proved. It is suggested that at sites where mineralization is to take place, the cells secrete alkaline phosphatase, which breaks down pyrophosphate ions, which would otherwise inhibit the growth of crystal surfaces. The supersaturated levels of calcium and phosphate ions in the tissue fluid (page 455) then enable all existing biological apatite crystal surfaces to grow spontaneously. 

CAT, chloramphenicol acetyltransferase gene LUC, luciferase gene SeAP, secreted alkaline phosphatase gene. 

COS-1, nuclear extracts containing receptor expressed in transfected COS-1 cells HL-60, cell extracts Baculovirus, recombinant receptors from the baculovirus/Sf21 expression system, wt, wild-type chimeric ER, RAR DEF or RXR D domain fused to ER DNA-binding domain CAT, chloramphenicol acetyl transferase gene LUC, luciferase gene SeAP, secreted alkaline phosphatase gene. 

Torisawa, Y.-S., Ohara, N., Nagamine, K., Kasai, S.. Yasukawa, T., Shiku, H., Matsue, T. Electrochemical monitoring of cellular signal transduction with a secreted alkaline phosphatase reporter system. Anal... 

HeLa cell transfection with various plasmid vectors encoding secreted alkaline phosphatase as the reporter protein is a widely applicable strategy. The plasmid vectors can be tailored to contain different responsive elements. For example, glucocorticoid, cAMP, the binding site of NFkB have been used in SECM single HeLa cell studies. ... 

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