A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity

Feinberg, Andrew P.; Vogelstein, Bert

doi:10.1016/0003-2697(83)90418-9

Cited by 23,152 publications

(8,576 citation statements)

References 20 publications

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“…The cDNA probes, a 0.7-kb fragment from the human IGF-IR sequence, and a 2.2-kb fragment of chicken ␤-actin were labeled with [␣-32 P]deoxycytidine triphosphate (DuPont New England Nuclear Research Products, Boston MA) using the random hexanucleotide primer method. 25 Northern blot hybridization was carried out in 5ϫ standard saline citrate (SSC), 5ϫ Denhardt's solution, 10% (wt/vol) dextran sulfate, 0.1% (wt/vol) sodium dodecyl sulfate (SDS), and 100 g/mL denatured salmon sperm DNA at 65°C for 18 hours. After hybridization, the membrane was washed for 15 minutes at room temperature with 2ϫ SSC followed by a final 15-minute wash in a solution consisting of 0.5ϫ SSC and 0.5% SDS (wt/vol) at 65°C.…”

Section: Northern Blot Analysismentioning

confidence: 99%

Treatment of human breast cancer cells with antisense RNA to the type I insulin-like growth factor receptor inhibits cell growth, suppresses tumorigenesis, alters the metastatic potential, and prolongs survival in vivo

Chernicky

Tan

et al. 2000

Cancer Gene Ther

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The type I insulin-like growth factor receptor (IGF-IR) plays an important role in the growth and transformation of breast cancer cells. In this study, we investigated the effects of treatment with an antisense IGF-IR construct on cells from the highly metastatic estrogen receptor-negative human breast cancer cell line MDA-MB-435s. The cells carrying the antisense IGF-IR had a markedly reduced expression of IGF-IR, had a significant decrease in cell proliferation, and lost the ability to form colonies in soft agar. There was a delay in tumor formation and a dramatic reduction in tumor size when cells carrying the antisense IGF-IR were injected into either nude or severe combined immunodeficient (scid) beige mice. We have also provided data that show that the scid beige mouse is a more suitable model for studying metastasis of the MDA-MB-435s cells. All of the scid beige mice injected with cells carrying the control construct had metastasis to the lungs, whereas lungs from the nude mice had no apparent metastatic sites after 11 weeks. When cells carrying antisense IGF-IR were injected subcutaneously in scid beige mice, the animals had a significant increase in survival compared with mice injected with cells carrying the control construct. Taken together, these results indicate that the IGF-IR can play a critical role in the progression of breast cancer. Our studies provide a basis for the development of future treatment strategies targeting the IGF-IR in metastatic breast cancer. Cancer Gene Therapy (2000) 7, 384 -395

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Section: Northern Blot Analysismentioning

confidence: 99%

Treatment of human breast cancer cells with antisense RNA to the type I insulin-like growth factor receptor inhibits cell growth, suppresses tumorigenesis, alters the metastatic potential, and prolongs survival in vivo

Chernicky

Tan

et al. 2000

Cancer Gene Ther

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“…The blots were then prehybridized and hybridized overnight with 32p-labelled cDNA probes. The cDNAs were radiolabelled by oligonucleotide priming [20]. Autoradiography of Northern blots was analyzed by densitometric scanning.…”

Section: Northern Analysis Of Mrna Levelsmentioning

confidence: 99%

The synthesis of human α‐2‐HS glycoprotein is down‐regulated by cytokines in hepatoma HepG2 cells

et al. 1988

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The regulation of the synthesis of a-2-HS glyeoprotein (AHSG) by inflammatory mediators from activated monocytes was studied on the human hepatoma cell line HepG2 and compared to that of albumin. Monocyte-conditioned medium, recombinant human interleukin-6 (rhlL6) and interleukin-lfl (rhlLlfl) all down-regulated the synthesis of AHSG. This decrease was found both at the protein and the mRNA level. The most efficient mediator was the monocyte-conditioned medium, when rhlLlfl was found to be less efficient than rhlL6. The combination of rhlL6 and rhlLlfl resulted in an additive down-regulation of the AHSG mRNA levels. Similar results were obtained with albumin. These data indicate that AHSG is a negative acute-phase protein whose synthesis is regulated by cytokines in a manner similar to that of albumin.Acute-phase protein; Glycoprotein ~-2-HS; Albumin; Interleukin-6; Interleukin-1

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“…The cDNA probe was labelled with [y"*P]dCTP using the random oligonucleotide priming method [15] to a specific activity of -2 x 10' dpm//g and further purified by spun column chromatography [16]. The labelled cDNA was hybridised at a concentration of 0.5 ng/ml to recombinant DNA on nitrocelluiose filters as described [9].…”

Section: Screeningmentioning

confidence: 99%

“…The entire PP-Y cDNA insert ( fig.2) [15] to 2 x 10' dpm//g specific activity. The probe was hybridised to polytene chromosome preparations obtained from the salivary gland of female Canton S strain D. melanogaster larvae as described [S] .…”

Section: In Situ Hybridisationmentioning

confidence: 99%

Molecular cloning and chromosomal localization of a novel Drosophila protein phosphatase

et al. 1989

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A 1.0 kilobase cDNA coding for the complete amino acid sequence of a putative protein phosphatase (314 amino acid residues, molecular mass 36 kDa) has been isolated from a Drosophila head cDNA library. The cDNA hybridises to a single site on the right arm of the second chromosome at cytological position 55Al-3. The deduced sequence of the protein, designated protein phosphatase-Y, is homologous to the catalytic subunits of Drosophila and rabbit protein phosphatase-la: (64 and 59% identity, respectively) and rabbit protein phosphatase-2A (39% identity). These and other comparisons demonstrate that this novel enzyme is not the Drosophila counterpart of mammalian protein phosphatases 1,2A, 2B, 2C or X.Protein phosphatase; cDNA cloning; Nucleotide sequence; Amino acid sequence; (Drosophila melanogaster)

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A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity

Cited by 23,152 publications

References 20 publications

Treatment of human breast cancer cells with antisense RNA to the type I insulin-like growth factor receptor inhibits cell growth, suppresses tumorigenesis, alters the metastatic potential, and prolongs survival in vivo

Treatment of human breast cancer cells with antisense RNA to the type I insulin-like growth factor receptor inhibits cell growth, suppresses tumorigenesis, alters the metastatic potential, and prolongs survival in vivo

The synthesis of human α‐2‐HS glycoprotein is down‐regulated by cytokines in hepatoma HepG2 cells

Molecular cloning and chromosomal localization of a novel Drosophila protein phosphatase

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