Millipore Sigma Vibrant Logo
Attention: We have moved. Merck Millipore products are no longer available for purchase on MerckMillipore.com.Learn More

407207 SignalBoost™Immunoreaction Enhancer Kit

407207
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

FormatDetection Methods
Liquid, Antibody DiluentChemiluminescence, colorimetric, or fluorescence

Products

Catalogue NumberPackaging Qty/Pack
407207-1KIT Plastic ampoule 1 kit
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
Western Blotting Tools
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.