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345818 RapidStep™ ECL Reagent

345818
  
Purchase on Sigma-Aldrich

Overview

Replacement Information
Description
Overview

This product has been discontinued.



A single component reagent for chemiluminescent detection of immobilized proteins, labeled with horseradish peroxidase, by immunoblotting. Provides a bright, stable signal with low signal:background. Does not require mixing of substrate and enhancer solutions. Volume is sufficient for covering up to 2000 cm2 of membrane.

Catalogue Number345818
Brand Family Calbiochem®
SynonymsChemiluminescent Substrate, Single Step
Application Data
Various amounts of mouse IgG, as indicated above, were dot blotted onto nitrocellulose. The protein was probed with a 1:5000 dilution of anti-mouse IgG HRP labeled antibody. RapidStep™ ECL Reagent is applied with a spray bottle as show below. Exposure time = 3 min.
OverviewRapidStep™ ECL is a 1 STEP chemiluminescent substrate reagent for detection of horseradish peroxidase (HRP) on immunoblots. It is a convenient, cost-effective, and ready-to-use reagent that provides a superior, longer-lasting signal than traditional ECL reagents.

RapidStep™ contains luminol and enhancer combined in one solution and is ready-to-use without mixing. A spray bottle is used to apply the solution directly onto the membrane.
References
Product Information
FormLiquid
Applications
Application NotesImmunoblotting
Application CommentsTo achieve an optimal signal:noise ratio, it is necessary to optimize the concentration of primary and secondary antibodies.
Protocol for Use
1. Remove the membrane containing immobilized proteins from the last wash and place on a flat surface, protein side facing up. Do not allow the membrane to dry
2. Spray RapidStep™ ECL Reagent on the surface of the membrane to sufficiently cover the entire membrane. Gently shake the container to ensure the membrane is evenly covered. Generally 2 sprays are sufficient to cover a 5 cm x 5 cm area.
3. Incubate the membrane for 1 min at room temperature without agitation. Proteins with low specificity or low reactivity with the primary antibody may require longer incubation times.
4. Drain the excess RapidStep™ ECL Reagent by holding the membrane vertically and touch the edge of the membrane against absorbent paper. Failure to drain sufficiently may lead to high background.
5. Wrap the membrane in plastic wrap and gently smooth out air pockets.
6. Place the membrane in a film cassette, protein side facing up.
7. Place X-ray film on top of the membrane, close the cassette and expose for an appropriate period of time. Optimal exposure times should be determined for each experiment.
Biological Information
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
R PhraseR: 36/38

Irritating to eyes and skin.
S PhraseS: 26-36-45

In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
Wear suitable protective clothing.
In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).
Product Usage Statements
Storage and Shipping Information
Ship Code Blue Ice Only
Toxicity Irritant
Storage +2°C to +8°C
Protect from Light Protect from light
Do not freeze Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
345818 0

Documentation

RapidStep™ ECL Reagent MSDS

Title

Safety Data Sheet (SDS) 

RapidStep™ ECL Reagent Certificates of Analysis

TitleLot Number
345818
Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision19-November-2009 RFH
SynonymsChemiluminescent Substrate, Single Step
ApplicationImmunoblotting
Application Data
Various amounts of mouse IgG, as indicated above, were dot blotted onto nitrocellulose. The protein was probed with a 1:5000 dilution of anti-mouse IgG HRP labeled antibody. RapidStep™ ECL Reagent is applied with a spray bottle as show below. Exposure time = 3 min.
DescriptionA single component reagent for chemiluminescent detection of immobilized proteins, labeled with horseradish peroxidase, by immunoblotting. Volume supplied is sufficient for covering up to 2000 cm2 of membrane.
FormLiquid
CommentsTo achieve an optimal signal:noise ratio, it is necessary to optimize the concentration of primary and secondary antibodies.
Protocol for Use
1. Remove the membrane containing immobilized proteins from the last wash and place on a flat surface, protein side facing up. Do not allow the membrane to dry
2. Spray RapidStep™ ECL Reagent on the surface of the membrane to sufficiently cover the entire membrane. Gently shake the container to ensure the membrane is evenly covered. Generally 2 sprays are sufficient to cover a 5 cm x 5 cm area.
3. Incubate the membrane for 1 min at room temperature without agitation. Proteins with low specificity or low reactivity with the primary antibody may require longer incubation times.
4. Drain the excess RapidStep™ ECL Reagent by holding the membrane vertically and touch the edge of the membrane against absorbent paper. Failure to drain sufficiently may lead to high background.
5. Wrap the membrane in plastic wrap and gently smooth out air pockets.
6. Place the membrane in a film cassette, protein side facing up.
7. Place X-ray film on top of the membrane, close the cassette and expose for an appropriate period of time. Optimal exposure times should be determined for each experiment.
Storage Protect from light
+2°C to +8°C
Do Not Freeze Ok to freeze
Toxicity Irritant