millipore,密理博,345818,RapidStep ECL Reagent

Description
Overview	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 Number	345818
Brand Family	Calbiochem®
Synonyms	Chemiluminescent 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.
Overview	RapidStep™ 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.

Product Information
Form	Liquid

Applications
Application Notes	Immunoblotting
Application Comments	To 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.

Applications

Application Notes

Immunoblotting

Application Comments

To 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.

Safety Information
R Phrase	R: 36/38 Irritating to eyes and skin.
S Phrase	S: 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).