Blot analysis

Fig. 1. Southern blot analysis of DNA showing (a) step 1, an agarose gel containing separated restriction fragments of DNA, denoted by (—), which is immersed in NaOH to denature the double-stranded stmcture of DNA, and then transferred by capillary flow to a nitrocellulose filter. In step 2, the bound DNA is allowed to hybridize to a labeled nucleic acid probe, and the unbound probe is washed off In step 3, the filter is placed into contact with x-ray film resulting in (b) bands of exposure on the film which are detected after development and correspond to regions where the restriction fragment is...

A. Western blot analysis of membrane fractions from P. vulgaris probed with a PGIP-specific antibody. 1) PGIP purified from P. vulgaris] 2) and 3) membranes 4) supernatant of the membrane preparation. 

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

Figure 4. RNA Blot Analysis of pSubunit, PG, and D21 Expression during Fruit Development and Ripening. Total RNA (25 pg) isolated from the indicated tomato tissues was probed with eiAer a psubunit cDNA clone, a cDNA for the catalytic PG polypeptide, or a cDNA for the constitutively expressed mRNA D21. Identical specific activities were used in each hybridization and all blots were exposed for 8 hr.

Further characterization by SDS-PAGE and Western blot analysis of the culture filtrate of a 6 day-old culture of lindemuthiaimm showed that only one endoPG of the same molecular weight (42 kDa) as the pure endoPG (Hugouvieux et al., 1995) was induced on pectin. 

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. 

The entire eluted fraction and the aliquots containing 3% of input and 3% of flow-through are then mixed with 4x sample loading buffer, boiled for 3 min at 95°, and subjected to Western blot analysis, as has been described in Section 2.4. 

Dubeau L, Chandler LA, Gralow JR, et al. Southern blot analysis of DNA extracted from formalin-fixed pathology specimens. Cancer Res. 1986 46 2964-2969. 

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