Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
SignalBoost™ 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 Number
407207
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 method
Chemiluminescence, colorimetric, or fluorescence
Form
20 Miniblots or 1600 cm²
Format
Liquid, Antibody Diluent
Kit contains
Solution 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 use
SignalBoost™ 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 Conditions
Upon 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 contains
Solution 1 for Primary Antibodies, Solution 2 for Secondary Antibodies, and a user protocol.
Upon 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 use
SignalBoost™ 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.
Background
SignalBoost™ 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 assay
The 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 provided
Solution 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 protocol
Dilute 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 Trademarks
Calbiochem® is a registered trademark of EMD Chemicals, Inc. Interactive Pathways™ and SignalBoost™ are trademarks of EMD Chemicals, Inc.
