polyclonal rabbit anti livin ab  (Novus Biologicals)

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: Relative livin mRNA expression levels in normal adrenal glands (NAG), adrenocortical adenoma (ACA) and adrenocortical carcinoma (ACC) in the entire cohort of patients, expressed in log scale ( A ) and in 19 paired samples of tumor and corresponding NAG, of which 6 ACC ( B ) and 13 ACA ( C ) ( p < 0.005 per trend). Statistical analysis by Kruskall-Wallis test. N.s.= p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Expressing

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: ( A ) 4× agarose gel for the expression of livin isoforms α and β at mRNA analyzed by RT-PCR in a subgroup of 4 paired samples of tumors (2 adrenocortical adenomas = ACA, and 2 adrenocortical carcinomas = ACC) and adjacent normal = NAG. Hela cells were used as positive control and β2-microglobulin was used as internal standard. ( B ) Quantitative analysis of agarose gel bands of all 15 paired samples of adrenocortical tumors (ACT) and corresponding NAG ( p = 0.3 per trend). Each bar in the histograms represents the mean of the ratio livin α or livin β signal to β2-microglobulin signal. ( C ) Western blot analysis of livin isoforms expression in three of four paired samples of tumors (2 ACA and 1 ACC) and adjacent NAG that are showed also at mRNA levels. Whole cell lysate SK-MEL 28 was used as positive control and β-actin was used as internal standard. ( D ) Quantitative analysis of WB bands of all 15 paired samples of tumors and corresponding NAG ( p < 0.0001 per trend). Each bar in the histograms represents the mean of the ratio livin α or livin β signal to β-actin signal. Statistical analysis by Kruskall-Wallis test and Mann-Whitney U test. N.s. = p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Agarose Gel Electrophoresis, Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Western Blot, MANN-WHITNEY

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: ( A ) Normal adrenal gland (NAG) (A.1–2) with adjacent adrenocortical carcinoma (ACC) (A 3-4) stained for H&E (A.1 and A.3) and livin (A.2 and A.4). A.1b, 2b and A.3b, 4b are 10× enlarged and detailed images of the 2× NAG and ACC, respectively. Arrows point to tumor infiltration in the adjacent NAG. TC: capsule, separating NAG and ACC. ( B ) Example of ACC with H-score 3 and negative nuclei stained for H&E (B.1) and livin (B.2) in comparison with ( C ) adrenocortical adenoma (ACA) characterized by H-score 2 and positive nuclei stained for H&E (C.1) and livin (C.2). The .1b, 2b and C.1b, C.1c are 20× enlarged and detailed images of the 10× ACC and ACA, respectively. Scale bar: 100 μm.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Staining, Comparison

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: Livin staining in normal adrenal gland ( A ). It is possible to recognize the three different zone of the adrenal gland: the zona glomerulosa (ZG), the zona fasciculata (ZF) and the zona reticularis (ZR). The Figures ( B ) and ( C ) are 20× enlarged and detailed images of the 10× ZG, ZF and ZR, respectively. Scale bar: 100 μm.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Staining

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: ( A ) Livin cytoplasmic protein expression evaluated as H-score in normal adrenal glands (NAG, n = 20), adrenocortical adenomas (ACA, n = 58) and adrenocortical carcinomas (ACC, n = 192) ( p = 0.01 per trend). ( B ) Livin cytoplasm/nuclear protein expression ratio evaluated as the ratio between H-score and nuclear score in NAG ( n = 20), ACA ( n = 58) and ACC ( n = 192) ( p < 0.0001 per trend). Statistical analysis by Kruskall-Wallis test. N.s. = p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Expressing

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: Relative livin mRNA levels ( A ) and CASP3 mRNA levels ( B ) evaluated by qRT-PCR at both 48 h and 72 h in NCI-H295R cells after livin transfection in comparison with those transfected with the empty vector (pCMV6). All qRT-PCR experiments were conducted in triplicates. (A) After 72 hours from transfection, the mRNA level of livin α and livin β were significantly increased in livin -NCI-H295R cells in comparison with those transfected with empty vector ( p = 0.04 and p = 0.02 for livin α and livin β , respectively). (B) Livin overexpression was associated with a significant decrease of CASP3 mRNA levels after 72 hours from transfection ( p = 0.015 ). ( C ) WB analysis showed clearly higher livin α and β protein levels and a light decrease of cleaved caspase 3 protein levels at both 48 and 72 hours. ( D ) Quantitative analysis of livin α, livin β and cleaved caspase-3 by WB in livin -NCI- H295R transfected cells in comparison with those transfected with the empty vector after 72 h. Livin α and livin β were significantly increased in livin -NCI-H295R cells in comparison with pCMV6-NCI-H295R ( p = 0.0028 and p = 0.008 for livin α and livin β, respectively). Cleaved caspase-3 was only slightly decreased in livin -NCI- H295R transfected cells ( p = n.s .). Each bar in the histograms represents the mean of the ratio between cleaved casp3 signal and α-tubulin signal. Each point represents the value of this ratio deriving from four separate experiments. ( E ) Immunofluorescence staining showed a decrease of cleaved caspase-3 protein levels (red color) in livin -NCI-H295R cells (green color) compared to pCMV6-NCI-H295R. Cell nuclei were visualized with 4#-6-diamidino-2-phenylindole (DAPI, blue color). Magnification 60×. Statistical analysis by unpaired t-test with Welch's correction. n.s.= p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Quantitative RT-PCR, Transfection, Comparison, Plasmid Preparation, Over Expression, Immunofluorescence, Staining