Northern blotting analysis

B. Northern blot analysis of total RNA extracted from hypocotyls of P. vulgaris and hybridised to a radioactive pg/p-specific probe. 

Northern blot analysis revealed that expression of NHE-1 is tissue-specific with highest transcript abundance in stomach and minimal levels in liver and skeletal muscle. Moreover, there is physiological evidence that certain segments of the nephron lack a basolateral Na /H" exchanger [76]. Using RT-PCR, Krapf and Solioz [77] observed that NHE-1 transcripts are not detectable in SI and S2 segments of superficial nephrons. 

Levels of RNA (usually specific mRNAs) in a cell can be measured by well-established techniques such as northern blot analysis or by PCR analysis. However, the recent advent of DNA microarray technology has converted the identification and measurement of specific mRNAs (or... 

Northern blot analysis has revealed that myeloperoxidase transcripts are only present in cells of the granulocytic lineage, and mRNA species of 3.0-... 

Traditionally, Northern blot analysis of isolated total RNA or mRNA has been used to determine differences in levels of mRNA between and among different animal groups, tissues or time-points. Standard protocols for performing Northern blot hybridization are available (Sambrook and Russell, 2001). Radioisotopic and colorimetric methods are available for probe labeling. The advantage of Northern blot analysis, especially... 

Figure 9. Some factor secreting mutants expresses CSF-1 transcripts. Northern blot analysis. 12mg of total RNA/lane were analyzed. The major transcript was a 4.0 kb CSF-1 message. Results for parental Myl D7 cells, stroma dependent subclones derived and stroma independent mutants are shown 1, 3i-l 2, 5i-l 3, 6i-4 4, 61-5 5, 31-2 6, 4i-l 7, 5i-3 8, 6i-2 9, 6i-3 10, 6i-17 11, 6i-18 12, 6i-19 13, 6i-20 14, 61-21 15, 6i-22 16, 6i-23 17, 6i-26. Those stroma independent mutants that are secretors are also indicated. Although additional CSF-1 specific splice variants could be detected at low levels in MS-5 and Myl-D7 mutants with high CSF-1 expression, no mutant/cell line specific splice product could be shown. However, the size of the splice product detected depended on the probe that was used for hybridization. Two additional messages (3.2 kb and 2.3 kb) were detected by hybridization with a full length CSF-1 cDNA and only one additional message (2.3kb) was detected by hybridization with a 3 fragment of the cDNA.

Figure 5.20 Northern blot analysis. (From Lane, S. et al., /. Biol. Chem., 276,48988-48996, 2001. With permission.)...

---