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  • Cadmium accumulation and metallothionein overexpression in internal spermatic vein of patients with varicocele. 19362335

    OBJECTIVES: To determine the possible molecular mechanism for the thickened wall in the internal spermatic vein (ISV) of patients with varicocele, we examined the cadmium (Cd) content and metallothionein (MT) expression in these diseased vessels. Previous studies have shown that Cd might play a role in the etiology of varicocele-associated infertility. MT, a metal-binding protein, protects against cell apoptosis during hypoxia. METHODS: The study group consisted of 20 patients with grade 3 left varicocele. The control group consisted of 15 volunteers with left-sided indirect inguinal hernia. Through a left inguinal incision, a 1-cm section of the ISV was resected from each patient to measure the Cd and MT levels. The results were analyzed using Student's t test. RESULTS: The Cd content in the ISV was 59.84 +/- 5.7 ng/g in the control group and 192.1 +/- 24.2 ng/g in the varicocele group. The relative intensity of the MT band was 40.52 +/- 3.74 in the control group and 78.26 +/- 5.61 in the varicocele group. MT expression was greater in the varicocele group than in the control group, and its deposition in the vascular endothelial layer was predominant using immunohistochemistry staining and confocal laser scanning. CONCLUSIONS: The results of the present study have demonstrated a greater accumulation of Cd in the ISV of the varicocele group than in the control group. The high Cd content and hypoxic conditions would induce overexpression of MT in the diseased vessels to protect the vascular cells from apoptosis. This might be a mechanism for the thickened wall of the ISV in patients with varicocele.
    Tipo de documento:
    Referencia
    Referencia del producto:
    AP124A
    Nombre del producto:
    Goat Anti-Mouse IgG Antibody, Alkaline Phosphatase conjugate
  • A role for huntington disease protein in dendritic RNA granules. 20185826

    Regulated transport and local translation of mRNA in neurons are critical for modulating synaptic strength, maintaining proper neural circuitry, and establishing long term memory. Neuronal RNA granules are ribonucleoprotein particles that serve to transport mRNA along microtubules and control local protein synthesis in response to synaptic activity. Studies suggest that neuronal RNA granules share similar structures and functions with somatic P-bodies. We recently reported that the Huntington disease protein huntingtin (Htt) associates with Argonaute (Ago) and localizes to cytoplasmic P-bodies, which serve as sites of mRNA storage, degradation, and small RNA-mediated gene silencing. Here we report that wild-type Htt associates with Ago2 and components of neuronal granules and co-traffics with mRNA in dendrites. Htt was found to co-localize with RNA containing the 3'-untranslated region sequence of known dendritically targeted mRNAs. Knockdown of Htt in neurons caused altered localization of mRNA. When tethered to a reporter construct, Htt down-regulated reporter gene expression in a manner dependent on Ago2, suggesting that Htt may function to repress translation of mRNAs during transport in neuronal granules.
    Tipo de documento:
    Referencia
    Referencia del producto:
    Múltiplo
    Nombre del producto:
    Múltiplo
  • Prognostic value of NDRG1 and SPARC protein expression in breast cancer patients. 20369286

    An increasing number of studies have shown altered expression of secreted protein acidic and rich in cysteine (SPARC) and N-myc down-regulated gene (NDRG1) in several malignancies, including breast carcinoma; however, the role of these potential biomarkers in tumor development and progression is controversial. In this study, NDRG1 and SPARC protein expression was evaluated by immunohistochemistry on tissue microarrays containing breast tumor specimens from patients with 10 years of follow-up. NDRG1 and SPARC protein expression was determined in 596 patients along with other prognostic markers, such as ER, PR, and HER2. The status of NDRG1 and SPARC protein expression was correlated with prognostic variables and patient clinical outcome. Immunostaining revealed that 272 of the 596 cases (45.6%) were positive for NDRG1 and 431 (72.3%) were positive for SPARC. Statistically significant differences were found between the presence of SPARC and NDRG1 protein expression and standard clinicopathological variables. Kaplan-Meier analysis showed that NDRG1 positivity was directly associated with shorter disease-free survival (DFS, P < 0.001) and overall survival (OS, P < 0.001). In contrast, patients expressing low levels of SPARC protein had worse DFS (P = 0.001) and OS (P = 0.001) compared to those expressing high levels. Combined analysis of the two markers indicated that DFS (P < 0.001) and OS rates (P < 0.001) were lowest for patients with NDRG1-positive and SPARC-negative tumors. Furthermore, NDRG1 over-expression and SPARC down-regulation correlated with poor prognosis in patients with luminal A or triple-negative subtype breast cancer. On multivariate analysis using a Cox proportional hazards model, NDRG1 and SPARC protein expression were independent prognostic factors for both DFS and OS of breast cancer patients. These data indicate that NDRG1 over-expression and SPARC down-regulation could play important roles in breast cancer progression and serve as useful biomarkers to better define breast cancer prognosis.
    Tipo de documento:
    Referencia
    Referencia del producto:
    AB1858
    Nombre del producto:
    Anti-Osteonectin Antibody
  • A subset of the histone H3 lysine 9 methyltransferases Suv39h1, G9a, GLP, and SETDB1 participate in a multimeric complex. 20129054

    Lysine 9 of histone 3 (H3K9) can be mono-, di-, or trimethylated, inducing distinct effects on gene expression and chromatin compaction. H3K9 methylation can be mediated by several histone methyltransferases (HKMTs) that possess mono-, di-, or trimethylation activities. Here we provide evidence that a subset of each of the main H3K9 HKMTs, G9a/KMT1C, GLP/KMT1D, SETDB1/KMT1E, and Suv39h1/KMT1A, coexist in the same megacomplex. Moreover, in Suv39h or G9a null cells, the remaining HKMTs are destabilized at the protein level, indicating that the integrity of these HKMTs is interdependent. The four HKMTs are recruited to major satellite repeats, a known Suv39h1 genomic target, but also to multiple G9a target genes. Moreover, we report a functional cooperation between the four H3K9 HKMTs in the regulation of known G9a target genes. Altogether, our data identify a H3K9 methylation multimeric complex.
    Tipo de documento:
    Referencia
    Referencia del producto:
    07-551
    Nombre del producto:
    Anti-G9a Antibody
  • Quantitative Detection of µ Opioid Receptor: Western Blot Analyses Using µ Opioid Receptor Knockout Mice. 21886594

    Increasing evidence suggests that µ opioid receptor (MOP) expression is altered during the development of and withdrawal from substance dependence. Although anti-MOP antibodies have been hypothesized to be useful for estimating MOP expression levels, inconsistent MOP molecular weights (MWs) have been reported in studies using anti-MOP antibodies. In the present study, we generated a new anti-MOP antibody (N38) against the 1-38 amino acid sequence of the mouse MOP N-terminus and conducted Western blot analysis with wildtype and MOP knockout brain lysates to determine the MWs of intrinsic MOP. The N38 antibody detected migrating bands with relative MWs of 60-67 kDa in the plasma membrane fraction isolated from wildtype brain, but not from the MOP knockout brain. These migrating bands exhibited semi-linear density in the range of 3-30 µg membrane proteins/lane. The N38 antibody may be useful for quantitatively detecting MOP.
    Tipo de documento:
    Referencia
    Referencia del producto:
    AB5511
    Nombre del producto:
    Anti-Opioid Receptor Antibody, µ, pain
  • Altered expression pattern of testican-1 mRNA after brain injury. 22199127

    Testican, a chondroitin/heparan sulfate proteoglycan, is primarily expressed in neurons of the adult and embryonic mouse brain, suggesting its role in normal and/or proliferation and differentiation processes of neurons. However, the role of testican in injured brain remains unclear. In the present study we investigated testican-1 mRNA expression pattern after cryo-injury of the brain. In situ hybridization histochemistry revealed that testican-1 mRNA is induced in the region surrounding the necrotic tissue. Time course study of testican-1 mRNA showed the highest level of signal intensity at 7 days after the injury. To determine which cell types express testican-1 mRNA, we performed in situ hybridization histochemistry combined with immunohistochemistry of several cell markers. Testican-1 mRNA signals were detected in the proximal reactive astrocytes, whereas the distribution pattern of testican-1 mRNA positive cells was different from those of mature oligodendrocytes and activated microglia. In addition, signals for testican-1 mRNA overlapped with those of FGF-2 mRNA, showing that these molecules are coexpressed in reactive astrocytes. These results suggest a possibility that testican-1 plays a permissive role for regenerating axons in reactive astrocytes after injury.
    Tipo de documento:
    Referencia
    Referencia del producto:
    MAB328
    Nombre del producto:
    Anti-Oligodendrocytes Antibody, clone CE-1
  • The FGF-1-specific single-chain antibody scFv1C9 effectively inhibits breast cancer tumour growth and metastasis. 25124967

    Immunotherapy mediated by recombinant antibodies is an effective therapeutic strategy for a variety of cancers. In a previous study, we demonstrated that the fibroblast growth factor 1 (FGF-1)-specific recombinant antibody scFv1C9 arrests the cell cycle at the G0/G1 transition by blocking the intracrine FGF-1 pathway in breast cancer cells. Here, we further show that the overexpression of scFv1C9 in MCF-7 and MDA-MB-231 breast cancer cells by lentiviral infection resulted in decreased tumourigenicity, tumour growth and lung metastasis through FGF-1 neutralization. We found that scFv1C9 resulted in the up-regulation of p21, which in turn inhibited the expression of CDK2 and blocked cell cycle progression. To explore the potential role of scFv1C9 in vivo, we delivered the gene into solid tumours by electroporation, which resulted in significant inhibition of tumour growth. In tumour tissue sections, immunohistochemical staining of the cellular proliferation marker Ki-67 and the microvessel marker CD31 showed a reduction in the proliferative index and microvessel density, respectively, upon expression of scFv1C9 compared with the appropriate controls. Thus, our data indicate a central role for scFv1C9 in blocking the intracrine pathway of FGF-1, therefore, scFv1C9 could be developed in an effective therapeutic for breast cancer.
    Tipo de documento:
    Referencia
    Referencia del producto:
    05-345
    Nombre del producto:
    Anti-p21/WAF1/Cip1 Antibody
  • Urothelial beta-3 adrenergic receptors in the rat bladder. 21046653

    To investigate the distribution of beta-3 adrenergic receptors (β(3)ARs) in the rat bladder and to examine the contribution of urothelial β(3)ARs to agonist-induced suppression of bladder reflexes and relaxation of smooth muscle.Bladder tissue was collected from 8- to 10-month old female SD rats. In some samples, the urothelium was surgically separated from the smooth muscle. The expression and localization of βAR mRNA and β(3)AR protein were determined using RT-PCR and immunohistochemistry. Contractile responses to the specific β(3)AR agonists TAK-677 and BRL37344 were measured in bladder strips with or without the urothelium. The contribution of urothelial β(3)ARs to the micturition reflex was assessed in continuous cystometry in urethane anesthetized rats using intravesical delivery of β(3)AR agonists.RT-PCR detected mRNA of all βARs in urothelium and smooth muscle. Immunostaining detected β(3)ARs throughout the urothelium, in the smooth muscle, myofibroblast-like cells, and in the peripheral nerves. Ovariectomy did not change the distribution of β(3)ARs in any bladder structure. Intravesical administration of TAK-677 and BRL37344 (1-5 × 10(-4) M) decreased voiding frequency and amplitude of bladder contractions. In bladder strips in vitro both β(3)AR agonists (10(-12) to 10(-4) M) relaxed the smooth muscle in a concentration-dependent manner to the same extent in strips with and without the urothelium.In addition to their presence in bladder smooth muscle, β(3)ARs are present in the urothelium where their activation may alter reflex voiding via release of factor(s) that act on non-myocyte structures including the afferent and/or efferent nerves to influence bladder contractility.
    Tipo de documento:
    Referencia
    Referencia del producto:
    AB15688
    Nombre del producto:
    Anti-Adrenergic β3 receptor Antibody
  • Monoclonal antibodies to native basement membranes reveal heterogeneous immunoreactivity patterns. 2684928

    In this paper we describe the development of basement membrane (BM) reactive monoclonal antibodies (MA), by immunization of mice with intact denuded BM. The MA raised against denuded amniotic BM (clones 1052, 1053 and 1065) showed heterogeneous staining patterns. MA 1052 and 1053 reacted with epithelial BM of the epidermis and epidermal adnexa and furthermore with the epithelial alveolar BM in the lung and the superficial part of the epithelial BM in the gastrointestinal tract. MA 1065 showed immunoreactivity with the epithelial BM of epidermis and epidermal adnexa and the epithelial BM of trachea and oesophagus, and furthermore pericellular staining of the basal keratinocytes and basal corneal epithelial cells. MA 1087, raised against human glomerular BM, showed immunoreactivity with all BM, except the central epithelial BM in the cornea. The precise localization of the target epitopes in the BM was investigated on chemically cleaved human skin. Reactivity for the MA occurred predominantly in the BM lamina adherent to the dermis, suggesting that the target epitopes reside in the lamina densa and/or lamina fibroreticularis. We furthermore examined the nature of the epitopes by preincubation of tissue sections with various enzymes prior to immunohistochemistry. The reactivity of the target epitopes was not affected by bacterial collagenase, but after various protease treatments the reactivity disappeared, suggesting that the epitopes are not localized on the triple helical part of collagenous proteins.
    Tipo de documento:
    Referencia
    Referencia del producto:
    MAB458
  • Development, characterization and use of monoclonal antibodies against sTRAIL: measurement of sTRAIL by ELISA. 11730847

    Two monoclonal antibodies against tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL), designated VI10E and III6F, have been generated. These antibodies were useful in flow cytometry analysis, immunohistochemistry, immunoprecipitation and in the development of an immunoassay for the detection of soluble TRAIL (sTRAIL)in biological samples. The immunoassay was based on two monoclonal antibodies against TRAIL. VI10E was used as the capture antibody and bound TRAIL was detected with anti-TRAIL from R&D Systems which was digoxigenin (DIG)-labeled. This enzyme-linked immunosorbent assay (ELISA) was specific for TRAIL since a panel of other cytokines did not affect the signal. The immunoassay was suitable for the detection of sTRAIL in human serum and plasma samples, cell culture supernatants and cell lysates. In a preliminary screening, it was found that serum samples from human immunodeficiency virus (HIV)-infected patients contained sTRAIL, and all these positive samples were found in the AIDS group. Using the immunoassay, it was found that phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) to produce significant amounts of sTRAIL, the levels of which increased with exposure time. Thus, the immunoassay for TRAIL presented here represents a useful tool for measuring sTRAIL in various biological samples. It will also permit studies of release mechanisms as well as possible functions of the soluble form of this molecule.
    Tipo de documento:
    Referencia
    Referencia del producto:
    MABF118
    Nombre del producto:
    Anti-sTRAIL Antibody, clone III6F