The mCK-5 Multiprobe RNase Protection Assay Kit Can Yield Erroneous Results for the Murine Chemokines IP-10 and MCP-1

2000 ◽  
Vol 286 (2) ◽  
pp. 193-197 ◽  
Author(s):  
Bruno Luckow ◽  
Holger Maier ◽  
Silvia Chilla ◽  
Guillermo Pérez de Lema
Keyword(s):  
Rnase Protection Assay ◽  
Protection Assay ◽  
Rnase Protection

A new quantitative solution hybridisation-RNase protection assay for APP and APLP2 mRNA

1996 ◽  
Vol 43 (1-2) ◽  
pp. 77-84 ◽  
Author(s):  
Janet A Johnston ◽  
Svante Norgren ◽  
Göran Annerén ◽  
Richard F Cowburn ◽  
Lars Lannfelt
Keyword(s):  
Rnase Protection Assay ◽  
Protection Assay ◽  
Rnase Protection

Abnormal myocyte Ca2+homeostasis in rabbits with pacing-induced heart failure

1998 ◽  
Vol 275 (4) ◽  
pp. H1441-H1448 ◽  
Author(s):  
Atsushi Yao ◽  
Zhi Su ◽  
Akihiko Nonaka ◽  
Iram Zubair ◽  
Kenneth W. Spitzer ◽  
...  
Keyword(s):  
Heart Failure ◽  
Ventricular Myocytes ◽  
Rnase Protection Assay ◽  
Protection Assay ◽  
Rnase Protection ◽  
Contraction Coupling ◽  
Intracellular Ca ◽  
Rapid Pacing ◽  
Sodium Binding ◽  
The Times

To determine whether there are abnormalities in myocyte excitation-contraction coupling and intracellular Ca2+concentration ([Ca2+]i) homeostasis in pacing-induced heart failure (PF), we measured L-type Ca2+ current ( I Ca,L) and Na+/Ca2+exchanger current ( I Na/Ca) with voltage clamp and measured intracellular Na+ concentration ([Na+]i) and [Ca2+]iwith the use of sodium-binding benzofuran isophthalate (SBFI) and fluo 3 in ventricular myocytes isolated from control and paced rabbits. The peak systolic and diastolic levels and the amplitude of electrically stimulated [Ca2+]itransients (0.25 Hz, extracellular Ca2+ concentration = 1.08 mM) were significantly less in PF myocytes. Also, there was prolongation of the times to peak and decline of [Ca2+]itransients. I Ca,Ldensity was markedly decreased in PF myocytes. I Na/Ca at −40 mV elicited by rapid exposure to 0 Na+ solution with a rapid solution switcher was significantly reduced in PF myocytes, suggesting that the function of the Na+/Ca2+exchanger is impaired in these myocytes. In PF myocytes the decline of the [Ca2+]itransient when the Na+/Ca2+exchanger was abruptly disabled was markedly prolonged compared with the decline in control myocytes, consistent with depressed sarcoplasmic reticulum (SR) Ca2+-ATPase function. RNase protection assay showed decreased levels of Na+/Ca2+exchanger and SR Ca2+-ATPase mRNA in PF hearts, consistent with the function studies. We conclude that the functions of L-type Ca2+channels, Na+/Ca2+exchanger, and SR Ca2+-ATPase are impaired in myocytes from rabbit hearts with failure induced by rapid pacing. These abnormalities result in reduced [Ca2+]itransients and systolic and diastolic dysfunction and appear to account for the abnormal ventricular function observed.

RNase Protection Assay

2019 ◽  
pp. 109-116
Author(s):  
Jianzhu Zhao ◽  
Jun Tang ◽  
Justin Elfman ◽  
Hui Li
Keyword(s):  
Rnase Protection Assay ◽  
Protection Assay ◽  
Rnase Protection

Gestational changes in fetal renal and hepatic angiotensinogen mRNA and protein

1998 ◽  
Vol 10 (5) ◽  
pp. 399 ◽  
Author(s):  
David Y. Zhang ◽  
Eugenie R. Lumbers ◽  
June J. Wu
Keyword(s):  
Fetal Liver ◽  
Rnase Protection Assay ◽  
Protein Product ◽  
Late Gestation ◽  
Protection Assay ◽  
Rnase Protection ◽  
Fetal Sheep ◽  
Protein Levels ◽  
Adult Kidney ◽  
Liver And Kidney

The aim of the study was to determine the amount of angiotensinogen expression and its protein product in fetal sheep liver and kidney in the last third of gestation. Angiotensinogen mRNA was measured by RNase protection assay and its protein levels were measured by radioimmunoassay. Levels were measured at 80, 95, 111, 125 and 139 days. Angiotensinogen mRNA was present in all fetal liver and kidney samples tested. The ratio of hepatic angiotensinogen mRNA/18 S rRNA increased by 100% (P<0.001) and angiotensinogen levels increased by 33% (P<0.001) in fetal sheep from 80 to 139 d. Over the same period the ratio of renal angiotensinogen mRNA/18 S rRNA increased by 170% (P<0.001) and renal angiotensinogen protein increased by 41% (P<0.001). The levels of angiotensinogen mRNA and its protein in the adult kidney were less than in kidneys of 139 d old fetuses (P<0.01). There was a direct relationship between levels of angiotensinogen mRNA and its protein in the liver (r = 0.53, P<0.01, n = 25) and in the kidney (r = 0.75, P<0.0001, n = 24). These findings demonstrate that there is a significant increase in both hepatic and renal angiotensinogen gene expression in the last third of gestation in the fetal sheep and that this increase is associated with an increase of angiotensinogen levels in both tissues. This increase in angiotensinogen in late gestation could influence the activity of both the intrarenal and circulating renin angiotensin systems.

Differential expression of the epidermal K1 and K10 keratin genes during mouse embryo development

1990 ◽  
Vol 68 (2) ◽  
pp. 448-453 ◽  
Author(s):  
Thérèse Ouellet ◽  
Marc Lussier ◽  
Feridoun Babaï ◽  
Line Lapointe ◽  
André Royal
Keyword(s):  
Histological Examination ◽  
Intermediate Filaments ◽  
Mouse Embryo ◽  
Rnase Protection Assay ◽  
Epidermal Differentiation ◽  
Protection Assay ◽  
Mouse Development ◽  
Rnase Protection ◽  
Stage Of Development ◽  
Mouse Embryo Development

Induction of genes coding for the K1 and K10 keratins during mouse development was studied by measuring the accumulation of their respective mRNAs in day 10 to 17 embryos using an RNase protection assay. Although these two keratins are coexpressed in the suprabasal layers of the epidermis, it was found that while K1 mRNA was detectable as soon as day 10, K10 mRNA was not detectable before day 12. The expression of these genes at this stage of development was not expected since they are specifically associated with keratinization, a process that does not begin before day 17 of gestation. Histological examination of the epidermis of day 10 to 17 embryos suggests that both genes are induced in cells committed to epidermal differentiation, after stratification has started but before the onset of keratinization. It was also found that the two mRNAs increased in abundance steadily and significantly until day 16 and that, in spite of the expectation that filaments should contain equivalent amounts of each subunit, K1 mRNA remained more abundant than K10 mRNA at all times including in adult epidermis. These observations indicate that the two genes are regulated independently during development.Key words: intermediate filaments, keratinization, differentiation, RNase protection, histology.

scholarly journals Effector Mechanisms of Protection againstPseudomonas aeruginosa Keratitis in Immunized Rats

2001 ◽  
Vol 69 (5) ◽  
pp. 3295-3304 ◽  
Author(s):  
A. Thakur ◽  
J. Kyd ◽  
M. Xue ◽  
M. D. P. Willcox ◽  
A. Cripps
Keyword(s):  
Molecular Mechanisms ◽  
Rnase Protection Assay ◽  
Interleukin 2 ◽  
Bacterial Count ◽  
Opportunistic Pathogen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sprague Dawley ◽  
Protection Assay ◽  
Rnase Protection ◽  
Sprague Dawley Rats

ABSTRACT Pseudomonas aeruginosa is an opportunistic pathogen which causes sight-threatening corneal infections in humans. The purpose of this study was to evaluate various immunization routes that may provide protection against Pseudomonas keratitis and to define the molecular mechanisms involved in the protection. Sprague-Dawley rats (10 to 12 weeks old) were immunized using paraformaldehyde-killed P. aeruginosa (strain 6206) via oral, nasal, and intra-Peyer's patch (IPP) routes followed by an ocular topical booster dose. Scratched corneas were challenged with an infective dose of P. aeruginosa. Following clinical examination, eyes were enucleated for histology, polymorphonuclear leukocyte (PMN) quantitation, bacterial count, enzyme-linked immunosorbent assay, and RNase protection assay. PMN infiltration was higher early (4 h) during the infection in immunized rats than in nonimmunized rats. Later during the infection, the number of PMNs diminished in immunized rats while in nonimmunized animals the number of PMNs continued to increase. Bacteria were cleared much faster from immunized groups than from the nonimmunized group, and the nasally immunized group had the most efficacious response among the immunized groups. Nasal and IPP immunization groups had increased cytokine expression of interleukin-2 (IL-2) and IL-5 and differed from each other for IL-6. All three immunized groups had significantly reduced IL-1β levels when compared with the nonimmunized rats and a significantly altered profile for CINC-1 expression. This study has shown that the route of immunization modulates the inflammatory response to ocular P. aeruginosa infection, thus affecting the severity of keratitis and adverse pathology, with nasal immunization being the most effective.

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