Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.

Shi, Shan‐Rong; Key, Marc E.; Kalra, Krishan L.

doi:10.1177/39.6.1709656

Cited by 2,335 publications

(1,188 citation statements)

References 11 publications

Supporting

Mentioning

1,148

Contrasting

Unclassified

Order By: Relevance

“…Antigen retrieval for p16 INK4a immunostaining was performed by incubating slides in 0.01 M citrate buffer (pH 6.0) for 40 min at 951C in a water bath (Pasha, Montone and Tomaszewski, 1995), sections for pRb immunostaining in 0.1 M sodium citrate buffer (pH 6.0) for 20 min in a microwave oven (Shi, Key and Kalra, 1991). For p16 INK4a immunostaining, mouse monoclonal antibody clone E6H4 (mtm laboratories, Heidelberg, Germany) was used.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Independent prognostic significance of cell cycle regulator proteins p16INK4a and pRb in advanced-stage ovarian carcinoma including optimally debulked patients: a translational research subprotocol of a randomised study of the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group

et al. 2007

View full text Add to dashboard Cite

The purpose of the study is to test the hypothesis that expression of cell cycle regulatory proteins p16 INK4a and pRb is significantly associated with prognosis in ovarian carcinomas. We performed immunohistochemical analysis of p16 INK4a and pRb expression and correlated with survival in a series of 300 patients with FIGO stage IIb-IV ovarian carcinoma which were enrolled in a randomized prospective trial evaluating two different platinum and paxlitaxel chemotherapy combinations after radical surgery. p16 INK4a negative tumours (17/300; 6%) had a significantly worse prognosis (univariate analysis, Po0.001; multivariate analysis: odds ratio 2.41, P ¼ 0.009). Among p16 INK4a -positive tumours (283 out of 300; 94%), survival was better for patients with intermediate expression as compared to low or high expression levels (P ¼ 0.001). High expression levels of pRb were associated with an incremental deterioration of prognosis (univariate analysis, P ¼ 0.004; multivariate analysis: odds ratio 2.98, P ¼ 0.002). This observation held also true in the subgroup of optimally debulked patients (n ¼ 82), in whom the most important established prognostic factor, postoperative residual tumour cannot be applied. In conclusion p16 INK4a and pRb are independent prognostic factors in advancedstage ovarian carcinomas after radical surgery and postoperative chemotherapy. High pRb expression is a significant prognosticator in optimally debulked patients and may hold potential for subgroup stratification in postoperative treatment.

show abstract

Section: Immunohistochemistrymentioning

confidence: 99%

Independent prognostic significance of cell cycle regulator proteins p16INK4a and pRb in advanced-stage ovarian carcinoma including optimally debulked patients: a translational research subprotocol of a randomised study of the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group

et al. 2007

View full text Add to dashboard Cite

show abstract

“…Sections analyzed for PCNA were microwaved 5 min for antigen retrieval. 18 Frozen tissues were used for evaluation of microvessel density (MVD) (CD31-positive structures). Tissues were sectioned (8 -10 m thick), mounted on positively charged Superfrost slides and air-dried for 30 min.…”

Section: Determination Of Proliferation Microvessel Density Cell Dementioning

confidence: 99%

Effect of the vascular endothelial growth factor receptor‐2 antibody DC101 plus gemcitabine on growth, metastasis and angiogenesis of human pancreatic cancer growing orthotopically in nude mice

Bruns

Shrader

Harbison

et al. 2002

Intl Journal of Cancer

134

View full text Add to dashboard Cite

Vascular endothelial growth factor (VEGF) is the major pro-angiogenic factor for most tumors. VEGF expression has been shown to be associated with a poor prognosis in human pancreatic cancer. The purpose of our study was to determine the effect of blockade of VEGF receptor-2 activity with or without gemcitabine on tumor growth and metastasis in an orthotopic model of human pancreatic cancer in nude mice. Therapy with gemcitabine or DC101, a VEGF receptor-2 antibody, resulted in a significant reduction of primary pancreatic tumor growth compared to untreated controls. The combination of DC101 and gemcitabine inhibited primary pancreatic tumor growth and lymphatic metastasis to a greater degree than either agent alone. Treatment with DC101 decreased vessel counts and increased the area of hypoxic tumor tissue compared to controls. Immunofluorescent double staining for apoptotic endothelial cells demonstrated a significant increase in the number apoptotic endothelial cells 24 days after initiation of therapy with DC101 plus gemcitabine. DC101 plus gemcitabine also increased tumor cell death and decreased tumor cell proliferation in pancreatic tumors. These findings indicate that blockade of VEGF receptor activation interferes with the survival of tumor endothelial cells, resulting in a reduction of primary pancreatic tumor growth in nude mice. Furthermore, the data demonstrate that anti-VEGF receptor-2 therapy potentiates the tumoricidal effect of gemcitabine in this model. Anti-VEGF receptor-2 therapy in combination with gemcitabine may be a novel therapeutic approach for advanced pancreatic cancer.

show abstract

“…Antigen retrieval immunohistochemical stain was used to identify the source of inducible NO synthase (iNOS) in lung cells [33,34]. …”

Section: Methodsmentioning

confidence: 99%

Niacinamide mitigated the acute lung injury induced by phorbol myristate acetate in isolated rat's lungs

et al. 2012

View full text Add to dashboard Cite

BackgroundPhorbol myristate acetate (PMA) is a strong neutrophil activator and has been used to induce acute lung injury (ALI). Niacinamide (NAC) is a compound of B complex. It exerts protective effects on the ALI caused by various challenges. The purpose was to evaluate the protective effects of niacinamide (NAC) on the PMA-induced ALI and associated changes.MethodsThe rat's lungs were isolated in situ and perfused with constant flow. A total of 60 isolated lungs were randomized into 6 groups to received Vehicle (DMSO 100 μg/g), PMA 4 μg/g (lung weight), cotreated with NAC 0, 100, 200 and 400 mg/g (lung weight). There were 10 isolated lungs in each group. We measured the lung weight and parameters related to ALI. The pulmonary arterial pressure and capillary filtration coefficient (Kfc) were determined in isolated lungs. ATP (adenotriphosphate) and PARP [poly(adenosine diphophate-ribose) polymerase] contents in lung tissues were detected. Real-time PCR was employed to display the expression of inducible and endothelial NO synthases (iNOS and eNOS). The neutrophil-derived mediators in lung perfusate were determined.ResultsPMA caused increases in lung weight parameters. This agent produced pulmonary hypertension and increased microvascular permeability. It resulted in decrease in ATP and increase in PARP. The expression of iNOS and eNOS was upregulated following PMA. PMA increased the neutrophil-derived mediators. Pathological examination revealed lung edema and hemorrhage with inflammatory cell infiltration. Immunohistochemical stain disclosed the presence of iNOS-positive cells in macrophages and endothelial cells. These pathophysiological and biochemical changes were diminished by NAC treatment. The NAC effects were dose-dependent.ConclusionsOur results suggest that neutrophil activation and release of neutrophil-derived mediators by PMA cause ALI and associated changes. NO production through the iNOS-producing cells plays a detrimental role in the PMA-induced lung injury. ATP is beneficial, while PARP plays a deteriorative effect on the PMA-induced ALI. NAC exerts protective effects on the inflammatory cascade leading to pulmonary injury. This B complex compound may be applied for clinical usage and therapeutic regimen.

show abstract

Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.

Cited by 2,335 publications

References 11 publications

Independent prognostic significance of cell cycle regulator proteins p16INK4a and pRb in advanced-stage ovarian carcinoma including optimally debulked patients: a translational research subprotocol of a randomised study of the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group

Independent prognostic significance of cell cycle regulator proteins p16INK4a and pRb in advanced-stage ovarian carcinoma including optimally debulked patients: a translational research subprotocol of a randomised study of the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group

Effect of the vascular endothelial growth factor receptor‐2 antibody DC101 plus gemcitabine on growth, metastasis and angiogenesis of human pancreatic cancer growing orthotopically in nude mice

Niacinamide mitigated the acute lung injury induced by phorbol myristate acetate in isolated rat's lungs

Contact Info

Product

Resources

About