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Immobilon-P+Membrane


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  • When using Immobilon-P in a slot blot unit, the membrane is sticking to the plastic part of the devi…

    The sticking of the membrane is due to the type of plastic used in the blotting unit.

    One way to address this issue is to silicanize the surface of the plastic. This can be done using a product like Sigmacote (Sigma # SL-2). [The coating should be tested on a noncritical area of the unit, such as a small area on the exterior, to make sure that the solution is compatible with the type of plastic used in the device. The solvent for the silicone is heptane.] Pour a small amount of Sigmacote onto a kimwipe and then wipe across the surface that will be in contact with the membrane. Let the solvent dry off completely, and then wipe the surface with a clean kimwipe to remove any residue. Wash the unit thoroughly in Milli-Q water, and then proceed with the binding.

    Another potential problem is that the unit may be closed too tightly. To address this issue, close the unit with just enough pressure to obtain a seal; additional pressure is of no benefit.
    문서 타입:
    FAQ
    카탈로그 번호:
    C3117
    제품명:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • I've completed my transfer to Immobilon-P, and now I see translucent areas on the membrane that corr…

    No. When protein is transferred to Immobilon-P, the membrane becomes hydrophilic where the protein is bound. If the membrane is dried, or even partially dried, and then wetted with an aqueous buffer, the regions with bound protein will appear translucent.
    문서 타입:
    FAQ
    카탈로그 번호:
    C3117
    제품명:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • I transferred my protein to Immobilon-P and I can't find it. I've stained the gel and the membrane …

    If the pI of your proteins are greater than the pH of the transfer buffer, the proteins will travel in the opposite direction. The protein probably transferred into the running buffer. If you suspect that your protein has a high pI, try CAPS buffer and/or put membrane on both sides of the gel.
    문서 타입:
    FAQ
    카탈로그 번호:
    C3117
    제품명:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • If I do a dot blot with Immobilon-P should I prewet the membrane?…

    A dot blot can be done in two ways, via vacuum filtration with a dot blot apparatus or through intrusion with a tuberculin syringe. With the first method, the membrane must first be rendered hydrophilic using methanol (this allows the protein to adsorb onto the membrane) followed by a quick soak in water prior to assembly in the apparatus. When using the second method it isn't necessary to prewet with methanol. The user simply holds the syringe directly against the membrane and lets the pressure exerted from the syringe intrude the solution ( and protein) onto the membrane.
    문서 타입:
    FAQ
    카탈로그 번호:
    C3117
    제품명:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • How should I store the Immobilon-P membrane?…

    Prior to use the Immobilon-P membrane should be stored at room temperature. After use the membrane may be stored by allowing the membrane to dry, wrap in plastic wrap and store at 4º C. For short term (overnight) storage the membrane may be stored wet by wrapping in plastic and refrigerating.
    문서 타입:
    FAQ
    카탈로그 번호:
    C3117
    제품명:
    Immobilon® Membranes, Sandwiches and Blotting Filter Paper

  • When performing an ELISpot assay should MultiScreen plates with Immobilon-P membrane (MAIP and MSIP)…

    Yes. Wetting the membrane with 15 µL alcohol followed by two to three washes with sterile PBS will allow the antibody more intimate contact with the membrane. Do not vacuum the alcohol through the underdrain, rather it should be flicked out. Once the membrane is pre-wet with alcohol, do not allow it to dry for the duration of the assay.
    문서 타입:
    FAQ
    카탈로그 번호:
    C9039
    제품명:
    Elispot MultiScreen®HTS Filter Plates and Antibody Pairs

  • Oncogene-induced senescence as an initial barrier in lymphoma development. 16079837

    Acute induction of oncogenic Ras provokes cellular senescence involving the retinoblastoma (Rb) pathway, but the tumour suppressive potential of senescence in vivo remains elusive. Recently, Rb-mediated silencing of growth-promoting genes by heterochromatin formation associated with methylation of histone H3 lysine 9 (H3K9me) was identified as a critical feature of cellular senescence, which may depend on the histone methyltransferase Suv39h1. Here we show that Emicro-N-Ras transgenic mice harbouring targeted heterozygous lesions at the Suv39h1, or the p53 locus for comparison, succumb to invasive T-cell lymphomas that lack expression of Suv39h1 or p53, respectively. By contrast, most N-Ras-transgenic wild-type ('control') animals develop a non-lymphoid neoplasia significantly later. Proliferation of primary lymphocytes is directly stalled by a Suv39h1-dependent, H3K9me-related senescent growth arrest in response to oncogenic Ras, thereby cancelling lymphomagenesis at an initial step. Suv39h1-deficient lymphoma cells grow rapidly but, unlike p53-deficient cells, remain highly susceptible to adriamycin-induced apoptosis. In contrast, only control, but not Suv39h1-deficient or p53-deficient, lymphomas senesce after drug therapy when apoptosis is blocked. These results identify H3K9me-mediated senescence as a novel Suv39h1-dependent tumour suppressor mechanism whose inactivation permits the formation of aggressive but apoptosis-competent lymphomas in response to oncogenic Ras.
    문서 타입:
    Reference
    카탈로그 번호:
    07-332
    제품명:
    Anti-HP1γ Antibody

  • Clinical and biochemical correlates of insoluble alpha-synuclein in dementia with Lewy bodies. 16482476

    Alpha-synuclein is a major constituent of Lewy bodies, the fibrillar aggregates that form within neurons in Parkinson's disease and dementia with Lewy bodies (DLB). Recent biochemical data show that alpha-synuclein accumulates in Parkinson's disease in a detergent insoluble form. We now examine the relationship between detergent insoluble alpha-synuclein and the presence of Lewy bodies, clinical measures of dementia and biochemical parameters in a series of individuals with DLB. We found that Triton X-100 insoluble alpha-synuclein enriched nearly twofold in the temporal cortex of patients with DLB compared to age-matched controls. By contrast the total amount of alpha-synuclein protein was unchanged. Surprisingly, the degree of Triton X-100 insoluble alpha-synuclein did not correlate with either the duration of illness or the number of Lewy bodies counted using stereological methods from an adjacent block of tissue. However, the Triton X-100 soluble fraction of alpha-synuclein did correlate strongly with the expression of several heat shock proteins (HSPs) in DLB but not control cases, suggesting a coordinated HSP response in DLB neocortex.
    문서 타입:
    Reference
    카탈로그 번호:
    Multiple
    제품명:
    Multiple