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407207 | SignalBoost™Immunoreaction Enhancer Kit

407207
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      Replacement Information

      Tabela com principais espec.

      FormatDetection Methods
      Liquid, Antibody Diluent Chemiluminescence, colorimetric, or fluorescence

      Preço e Disponibilidade

      Número de catálogoDisponibilidade Embalagem Qtde/Emb. Preço Quantidade
      US1407207-1KIT
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          Frasco de vidro 1 kit
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          Description
          OverviewSignalBoost™ Immunoreaction Enhancer Kit is designed to enhance the desired signal in assays such as immunoblotting, dot blotting, and ELISA. This kit significantly improves sensitivity and discrimination when assessing protein targets of low immunoreactivity or low expression levels. Enhancement of signal is especially marked for low affinity antibodies. It replaces conventional antibody diluents with Solution 1, which is used to dilute primary antibodies and Solution 2, which is used to dilute secondary antibodies. No additional steps, materials, or increase of assay time is required.

          Increases signal: minimizes background resulting in high signal to noise ratios
          Versatile: can be used with nitrocellulose or PVDF membranes and is compatible with chemiluminescent or colorimetric detection systems
          Convenient: ready-to-use format, no dilution necessary
          Catalogue Number407207
          Brand Family Calbiochem®
          Application Data
          Detection of protein kinase C by immunoblotting. Samples: whole cell lysate from K562 cells. Immunoblotting conditions: nitrocellulose membrane blocked overnight with 5% milk in PBST; all washes were performed with PBST. Primary antibody: Anti-PKC (Ab-2) Mouse mAb (MC5) (Cat. No. OP74) diluted 1:1000 in 5% milk/PBST (lane A) or in Solution 1 (lane B) (incubation for 1 h ). Secondary antibody: Anti-Mouse IgG diluted 1:10,000 in 5% milk/PBST (lane A) or Solution 2 (lane B). Detection: chemiluminescence (30 s exposure).

          ELISA results generated with and without dilution of antigen and detection antibody using SignalBoost™ reagent.
          Materials Required but Not Delivered Any equipment necessary for performing the desired immunoassay
          Primary antibody
          Secondary antibody (labeled with HRP, AP, etc.) specific to the primary antibody
          References
          Product Information
          Detection methodChemiluminescence, colorimetric, or fluorescence
          Form20 Miniblots or 1600 cm²
          FormatLiquid, Antibody Diluent
          Kit containsSolution 1 for Primary Antibodies, Solution 2 for Secondary Antibodies, and a user protocol.
          Applications
          Biological Information
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          Product Usage Statements
          Intended useSignalBoost™ Immunoreaction Enhancer Solution is a reactive reagent for use in immunoassays, and was developed to resolve the problems of low sensitivity and high background often encountered during procedures such as immunoblotting and ELISA (enzyme-linked immunosorbent assay). It can be used in a range of immunoassays.
          Storage and Shipping Information
          Ship Code Blue Ice Only
          Toxicity Standard Handling
          Storage +2°C to +8°C
          Storage ConditionsUpon receipt, store the entire contents of the kit at 4°C. Protect from light. Following initial use, this product is stable for up to 1 year at 4°C.
          Protect from Light Protect from light
          Do not freeze Ok to freeze
          Packaging Information
          Transport Information
          Supplemental Information
          Kit containsSolution 1 for Primary Antibodies, Solution 2 for Secondary Antibodies, and a user protocol.
          Specifications

          Documentation

          Certificados de análise

          TítuloNúmero do lote
          407207

          Brochura

          Título
          Western Blotting Tools
          Protocolo do usuário

          Revision02-May-2011 JSW
          Form20 Miniblots or 1600 cm²
          FormatLiquid, Antibody Diluent
          Detection methodChemiluminescence, colorimetric, or fluorescence
          StorageUpon receipt, store the entire contents of the kit at 4°C. Protect from light. Following initial use, this product is stable for up to 1 year at 4°C.
          Intended useSignalBoost™ Immunoreaction Enhancer Solution is a reactive reagent for use in immunoassays, and was developed to resolve the problems of low sensitivity and high background often encountered during procedures such as immunoblotting and ELISA (enzyme-linked immunosorbent assay). It can be used in a range of immunoassays.
          BackgroundSignalBoost™ enhances antigen-antibody reactions, yielding signals several times to several tens of times higher than those obtained with conventional methods. It has been verified that enhancement is particularly marked for relatively inactive antibodies. Compositions optimized for primary and secondary antibodies respectively minimize background and yield high S/N ratios. It can be used for various assays involving antigen-antibody reactions, such as immunoblotting, dot blotting and ELISA. Components do not affect the activity of enzymes used for labeling (peroxidase, alkaline phosphatase, etc.), allowing use in assays using any of these enzymes. It can also be used for detection of both fluorescence and coloration. For convenience, SignalBoost™ is supplied in a ready-to-use form, is at an appropriate concentration for use, and does not need to be diluted. Simply dilute antibodies with this product instead of conventional diluents to use. It does not lengthen the time required for the assay.
          Principles of the assayThe SignalBoost™ Immunoreaction Enhancer Kit provides a set of reagents for the dilution of primary and secondary antibodies. Solution 1 for Primary Antibodies is used to dilute primary antibodies to the recommended concentration, while Solution 2 is used to dilute secondary antibodies to the recommended concentration. Both solutions are designed to replace other diluents, such as TBS, TBS-T, PBS, PBS-T, etc. The remainder of the immunassay is carried out per standard protocols.
          Materials providedSolution 1 for Primary Antibodies (Kit Component No. KP31812-200ML): 1 bottle, 200 ml
          Solution 2 for Secondary Antibodies (Kit Component No. KP31855-200ML): 1 bottle, 200 ml
          Materials Required but not provided Any equipment necessary for performing the desired immunoassay
          Primary antibody
          Secondary antibody (labeled with HRP, AP, etc.) specific to the primary antibody
          Precautions and recommendations When using SignalBoost™ Immunoreaction Enhancer solutions with only one antibody, Solution 2 for Secondary Antibodies should be used.
          For sandwich ELISA involving solid-phase antibody and antigen, Solution 1 for Primary Antibodies may be used to dilute the antigen.
          This product may be used in combination with blocking agents such as casein, BSA, etc. However, since it contains substances with blocking activity, the concentration of blocking agents can become excessively high if used in combination.
          This product cannot be used for blocking reactions. It cannot be used for solidification of antigen or antibody during ELISA.
          Detailed protocolDilute primary antibodies to the recommended concentration using Solution 1 for Primary Antibodies. The dilution factor will depend on the antibody; refer to the manufacturer's guidelines for dilution. When using for ELISA detection, it is also recommended that the antigen also be diluted with Solution 1 prior to adding the samples to the wells of the ELISA plate. Dilute secondary antibodies to the recommended concentration using Solution 2 for Secondary Antibodies. The dilution factor will depend on the antibody; refer to the manufacturer's guidelines for dilution. Proceed with all immunoassays using standard protocols.
          Assay characteristics and examples

          Figure 1: Immunoblot Analysis

          Detection of protein kinase C by immunoblotting. Samples: whole cell lysate from K562 cells. Immunoblotting conditions: nitrocellulose membrane blocked overnight with 5% milk in PBST; all washes were performed with PBST. Primary antibody: Anti-PKC (Ab-2) Mouse mAb (MC5) (Cat. No. OP74) diluted 1:1000 in 5% milk/PBST (lane A) or in Solution 1 (lane B) (incubation for 1 h ). Secondary antibody: Anti-Mouse IgG diluted 1:10,000 in 5% milk/PBST (lane A) or Solution 2 (lane B). Detection: chemiluminescence (30 s exposure).


          Figure 2: ELISA Analysis

          ELISA results generated with and without dilution of antigen and detection antibody using SignalBoost™ reagent.

          Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
          Interactive Pathways™ and SignalBoost™ are trademarks of EMD Chemicals, Inc.