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  • Phosphatidylinositol 3-kinase/AKT signalling pathway components in human breast cancer: clinicopathological correlations. 17649772

    BACKGROUND: The phosphatidylinositol 3-kinase (PI3K)/Akt signalling pathway plays a major role in the regulation of breast cancer growth and survival, but the clinical value of its components in human tumours is unclear. PATIENTS AND METHODS: PI3K was analysed using Western blotting with monoclonal antibodies to the p85 subunit in tumour and adjacent mammary gland samples from 33 breast cancer patients. Activated Akt1 (pAkt1) expression was quantified in 46 sample pairs by a direct sandwich ELISA assay. RESULTS: Tumour PI3K expression was increased in 79% of the investigated sample pairs and was not associated with the main clinico-pathological features. Only 49% of breast cancers had increased pAkt1, but the frequency of its elevation was positively associated with tumour size and histological grade, and controversially related to estrogen and progesterone receptor status. CONCLUSION: Increased PI3K but not pAkt1, expression appears to be a widespread feature of human breast cancer indicating the different roles of the two components of one signalling system.
    Document Type:
    Reference
    Product Catalog Number:
    05-591
    Product Catalog Name:
    Anti-Akt/PKB Antibody, PH Domain, clone SKB1

  • A novel detection method of cleaved plasma high-molecular-weight kininogen reveals its correlation with Alzheimer's pathology and cognitive impairment. 30310850

    Accumulation of β-amyloid is a pathological hallmark of Alzheimer's disease (AD). β-Amyloid activates the plasma contact system leading to kallikrein-mediated cleavage of intact high-molecular-weight kininogen (HKi) to cleaved high-molecular-weight kininogen (HKc). Increased HKi cleavage is observed in plasma of AD patients and mouse models by Western blot. For potential diagnostic purposes, a more quantitative method that can measure HKc levels in plasma with high sensitivity and specificity is needed.HKi/c, HKi, and HKc monoclonal antibodies were screened from hybridomas using direct ELISA with a fluorescent substrate.We generated monoclonal antibodies recognizing HKi or HKc specifically and developed sandwich ELISAs that can quantitatively detect HKi and HKc levels in human. These new assays show that decreased HKi and increased HKc levels in AD plasma correlate with dementia and neuritic plaque scores.High levels of plasma HKc could be used as an innovative biomarker for AD.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple
    Product Catalog Name:
    Multiple

  • Development of a specific ELISA to measure BACE1 levels in human tissues. 21889954

    The aspartyl protease BACE1 is the rate limiting enzyme in the synthesis of amyloid beta, which accumulation in the human brain is a hallmark of Alzheimer's disease (AD). BACE1 has been proposed as a surrogate marker of AD; however, very few BACE1 immunoassays have been reported. In the present study we have screened ten BACE1 antibodies by Western blot and several antibody pairs to develop a new BACE1 sandwich ELISA procedure. We identified one pair that showed little background and good reproducibility. Several dilution buffers and sample denaturation methods were tried to partially unfold BACE1 before capture. We found that dilution in PBS followed by 10 min incubation at 50°C critically improves the performance of the assay. Finally, we successfully measured BACE1 levels in a few human brain and platelet lysates as well as in plasma and AD CSF. We anticipate that this assay will lay the ground to accurately measure BACE1 levels in human tissues, which could facilitate the molecular diagnosis of AD in the near future.
    Document Type:
    Reference
    Product Catalog Number:
    MAB5308
    Product Catalog Name:
    Anti-BACE Antibody, CT, clone 61-3E7

  • A monoclonal-based, two-site enzyme immunoassay of human insulin. 3553334

    A procedure is described for the efficient production of insulin-specific monoclonal antibodies, which involves primary and secondary immunization of BALB/c mice in the hind footpads with bovine or porcine insulin and fusion of lymphocytes from popliteal lymph nodes with a P3x63 murine myeloma line. With this protocol, over 200 positive hybrids were obtained from four separate fusions. Dissociation constants of 31 purified monoclonals, cross-reacting with human insulin, were determined by two different methods and ranged between 4 X 10(-10) and 2 X 10(-6) mol/l. 24 monoclonals were biotinylated, paired in all possible combinations and tested by ELISA for their capacity to simultaneously bind to human insulin in a two-site assay. More than 40 monoclonal pairs were found which formed a sandwich with the hormone. The development of a simple and rapid one-step enzyme immunoassay is described, which involves a first monoclonal bound to the wells of a microtiter plate and a second monoclonal conjugated to alkaline phosphatase. With this assay, insulin can be determined in a range between 0.08 and 7.5 ng/ml in 3-4 h.
    Document Type:
    Reference
    Product Catalog Number:
    MAB1697

  • Total and high molecular weight adiponectin in patients with coronary artery disease. 19430341

    BACKGROUND: Serum adiponectin inversely correlates with the extent of coronary artery disease (CAD). OBJECTIVES: To investigate the clinical significance of measuring high molecular weight (HMW) adiponectin in addition to total adiponectin from different samples (peripheral veins and ostia of coronary arteries) in patients with CAD. METHODS: We studied 134 patients; 57 with acute coronary syndrome (ACS), 44 with stable angina and 33 healthy patients. Total and HMW adiponectin were measured in the coronary ostia and peripheral veins simultaneously. Venous levels of lipid profile, C-reactive protein, tumour necrosis factor-alpha, interleukin-6 and insulin were measured. RESULTS: Mean levels of HMW adiponectin in the coronary ostia were significantly correlated with the venous levels of total (r = 0.33, P = 0.002) and HMW (r = 0.37, P = 0.002) adiponectin. Mean levels of total and HMW adiponectin were lower in CAD patients versus controls (6.9 +/- 0.5 versus 7.9 +/- 0.7 mug/ml, P = 0.3 and 1.9 +/- 0.2 versus 3.1 +/- 0.3 mug/ml, P = 0.003). In the coronary ostia, levels of HMW adiponectin were higher in ACS than those with stable angina (1.1 +/- 0.1 versus 0.8 +/- 0.1 mug/ml, P = 0.2). In patients with CAD, levels of ostial adiponectin (total and HMW) were significantly lower in diabetic than non-diabetic patients (6.5 +/- 0.8 versus 9.4 +/- 1.1 mug/ml, P = 0.04 and 0.8 +/- 0.12 versus 1.2 +/- 0.1 mug/ml, P = 0.03). Mean levels of venous adiponectin (total and HMW) were non-significantly lower in diabetic patients (5.9 +/- 0.7 versus 7.7 +/- 0.7, P = 0.12 and 1.8 +/- 0.4 versus 1.9 +/- 0.1, P = 0.8). CONCLUSION: Measurement of HMW adiponectin and its ratio to total adiponectin may be a better marker for CAD than total adiponectin levels. Ostial levels of adiponectin in ACS may indicate re-distribution of adiponectin molecules towards the acute lesions. The low HMW adiponectin levels in diabetes mellitus may in part explain the worse outcome of CAD in diabetics.
    Document Type:
    Reference
    Product Catalog Number:
    EZHI-14K
    Product Catalog Name:
    Human Insulin ELISA

  • Decreased levels of brain-derived neurotrophic factor in the remitted state of unipolar depressive disorder. 22292883

    Hasselbalch BJ, Knorr U, Bennike B, Hasselbalch SG, Greisen Søndergaard MH, Vedel Kessing L. Decreased levels of brain-derived neurotrophic factor in the remitted state of unipolar depressive disorder. Objective:  Decreased levels of peripheral brain-derived neurotrophic factor (BDNF) have been associated with depression. It is uncertain whether abnormally low levels of BDNF in blood are present beyond the depressive state and whether levels of BDNF are associated with the course of clinical illness. Method:  Whole-blood BDNF levels were measured in blood samples from patients with unipolar disorder in a sustained state of clinical remission and in a healthy control group. Participants were recruited via Danish registers, a method that benefits from the opportunity to obtain well-matched community-based samples as well as providing a high diagnostic validity of the patient sample. Results:  A total of 85 patients and 50 controls were included in the study. In multiple linear regression analyses, including the covariates age, gender, 17-item Hamilton Depression Rating Scale scores, body-mass index, education, smoking and physical exercise, patients with unipolar depressive disorder had decreased levels of BDNF compared to healthy control individuals [B = -7.4, 95% CI (-11.2, -3.7),  = 0.21 P < 0.001]. No association between course of clinical illness and BDNF levels was present. Conclusion:  Whole-blood BDNF levels seem to be decreased in patients remitted from unipolar depressive disorder, suggesting that neurotrophic changes may exist beyond the depressive state.
    Document Type:
    Reference
    Product Catalog Number:
    CYT306
    Product Catalog Name:
    ChemiKine Brain Derived Neurotrophic Factor, Sandwich ELISA
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