407207 SignalBoost™Immunoreaction Enhancer Kit

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407207
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Overview

Replacement Information

Key Specifications Table

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

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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

      SignalBoost™Immunoreaction Enhancer Kit SDS

      Title

      Safety Data Sheet (SDS) 

      SignalBoost™Immunoreaction Enhancer Kit Certificates of Analysis

      TitleLot Number
      407207

      Brochure

      Title
      SNAP i.d. 2.0 System Brochure

      Technical Info

      Title
      Western Blotting Quick Reference Product Guide
      User Protocol

      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.