Synthesis and Purification of Digoxigenin-Labeled RNA Probes for In Situ Hybridization

2007 ◽  
Vol 2007 (8) ◽  
pp. pdb.prot4778-pdb.prot4778 ◽  
Author(s):  
H. L. Sive ◽  
R. M. Grainger ◽  
R. M. Harland
Keyword(s):  
In Situ Hybridization ◽  
Rna Probes ◽  
Labeled Rna

In Situ Hybridization with Riboprobes: An Overview for Veterinary Pathologists

1998 ◽  
Vol 35 (3) ◽  
pp. 159-167 ◽  
Author(s):  
C. Brown
Keyword(s):  
In Situ Hybridization ◽  
Molecular Biology ◽  
Infectious Agents ◽  
Probe Target ◽  
Cellular Genes ◽  
Rna Probes ◽  
Pcr Technology ◽  
Labeled Rna ◽  
Target Combination

In situ hybridization using nonradioactively-labeled RNA probes is a technique that combines understanding of basic molecular biology and histopathologic interpretation. Recombinant or PCR technology can be used to produce probes that hybridize with a wide variety of cellular genes and infectious agents. Hybridization conditions can be optimized for each probe/target combination.

scholarly journals A fixative suitable for in situ hybridization histochemistry.

1993 ◽  
Vol 41 (6) ◽  
pp. 947-953 ◽  
Author(s):  
F Uehara ◽  
N Ohba ◽  
Y Nakashima ◽  
T Yanagita ◽  
M Ozawa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Proteinase K ◽  
Optimal Time ◽  
Hybridization Histochemistry ◽  
Tissue Sections ◽  
In Situ Hybridization Histochemistry ◽  
Rna Probes ◽  
Labeled Rna ◽  
The Stability

We compared the morphology and stability of hybridization signals between paraffin sections of rat retina fixed with commonly used 4% paraformaldehyde/PBS and those fixed with a fixative containing glutaraldehyde in in situ hybridization histochemistry, using a digoxigenin-labeled RNA probe complementary for beta-galactoside alpha 2,6-sialyltransferase mRNA. Retinal detachment was frequently observed in the sections fixed with 4% paraformaldehyde-PBS, whereas the morphology was satisfactorily preserved in those fixed with either 0.5% glutaraldehyde, 4% paraformaldehyde-PBS, or 2.5% glutaraldehyde-PBS. Without glutaraldehyde, it was difficult to determine the most appropriate length of proteinase K digestion of tissue sections for facilitating probe penetration, since the optimal time for definite hybridization was variable among the retinal cells in heterogeneous layers. By addition of glutaraldehyde to paraformaldehyde or with glutaraldehyde alone, it was easy to establish the appropriate time for the unmasking procedure, since intense mRNA signals were constant throughout the retina by proteinase K digestion for more than 30-40 min. Using a fixative that causes stronger cross-linking (e.g., glutaraldehyde) is recommended to improve not only the morphology but also the stability of hybridization signals in in situ hybridization histochemistry with paraffin embedding and digoxigenin-labeled RNA probes.

scholarly journals 2. Histocytochemistry Series In Situ Hybridization Using Non-radioisotope labeled RNA Probes (4)

2010 ◽  
Vol 6 (1) ◽  
pp. 23-29
Author(s):  
Takuya Mishima ◽  
Shan-shun Luo ◽  
Toshiyuki Takeshita ◽  
Toshihiro Takizawa
Keyword(s):  
In Situ Hybridization ◽  
Rna Probes ◽  
Labeled Rna

In Situ Hybridization of Labeled RNA Probes to Fixed Parhyale hawaiensis Embryos

2009 ◽  
Vol 2009 (1) ◽  
pp. pdb.prot5130-pdb.prot5130 ◽  
Author(s):  
E. J. Rehm ◽  
R. L. Hannibal ◽  
R. C. Chaw ◽  
M. A. Vargas-Vila ◽  
N. H. Patel
Keyword(s):  
In Situ Hybridization ◽  
Rna Probes ◽  
Parhyale Hawaiensis ◽  
Labeled Rna

scholarly journals Localization of mRNAs of two androgen-dependent proteins, SMR1 and SMR2, by in situ hybridization reveals sexual differences in acinar cells of rat submandibular gland.

1993 ◽  
Vol 41 (11) ◽  
pp. 1645-1649 ◽  
Author(s):  
I Rosinski-Chupin ◽  
C Rougeot ◽  
Y Courty ◽  
F Rougeon
Keyword(s):  
In Situ Hybridization ◽  
Submandibular Gland ◽  
Acinar Cells ◽  
Sexual Differences ◽  
Mrna Levels ◽  
Rna Probes ◽  
Labeled Rna ◽  
Rat Submandibular Gland ◽  
Convoluted Tubules

Androgen-dependent sexual differences in the granular convoluted tubules of mouse and rat submandibular glands (SMG) have been extensively reported. We studied two major androgen-dependent mRNAs of the rat SMG encoding proteins named SMR1 and SMR2. To determine which cell type in the SMG is responsible for synthesis of these mRNAs, we performed in situ hybridization with digoxigenin-labeled RNA probes coupled with alkaline phosphatase detection. We show that SMR1 and SMR2 mRNAs are synthesized in the acinar cells of the SMG. A clear difference in SMR1 and SMR2 mRNA levels in male and female is demonstrated. During the course of this study we also confirmed the acinar localization of mRNAs encoding the glutamine/glutamic acid-rich proteins (GRP) of rat SMG. Our data are the first clear evidence of androgen-dependent sexual differences in acinar cells of rat submandibular gland.

Sign in / Sign up

Export Citation Format

Share Document