cat # sl100309 store at 4 0 c

Cat # SL100309 Store at 4 0 C LumiGOLD™ ECL Western Blot ting Detection Kit Standard Pack 250 ml Large Pack 1000 m l 15875 Gaither Drive Gaithersburg, MD 20877 FAX. 301-560-4919 TEL. 301-330-5966 Toll Free. 1-866-918-6812 Email: [email protected] Web: www.signagenlabs.com Description: LumiGOLD™ ECL Kit is a complete kit with ready-to-u se reagents for chemiluminescent detection of immob ilized proteins (Western blotting) or immobilized n ucleic acids (Southern or Northern), conjugated wit h HRP directly or indirectly. The use of enhanced chemiluminescence was introduce d by Thorpe and Kricka (1,2). In the presence of hy drogen peroxide (H 2 O 2 ), Horseradish peroxidase (HR P) catalyzes the oxidation of cyclic diacylhydrazid es, such as luminol. Immediately following the oxid ation, the luminol is in an excited state (intermediate reaction product), w hich decays to the ground state by emitting light. Strong enhancement of the light emission is produce d by enhancers, such as phenolic compounds. Using t his method, it is possible to detect membrane immob ilized specific antigens, or sequences of nucleic a cids, labeled directly with HRP or indirectly with HRP-labeled antibodies/streptavidin. Quick Protocol: 1. Mix an equal volume of LumiGOLD™ ECL Kit Reagent A and Reagent B to give sufficient solution to cove r the membrane (0.1ml / cm2). Let the detection mi x equilibrate for 2 minute at room temperature. 2. Drain the excess buffer from the washed blots. D o not let the membrane dry out. Add the detection mix directly to the blot (protein side up). Incubat e for 2 minute at room temperature. 3. Drain off excess detection mix and wrap the memb rane in saran wrap. Gently remove air pockets. 4. Place the blots immediately , protein side up, in the film cassette. Switch off the lights and use re d safety light. Place a sheet of film on the blot, close the cassette and expose for 30-90 seconds or longer. 5. Replace the exposed film with a new one, close t he cassette and develop the first exposed film. 6. Expose the second film for a suitable time accor ding to the signal intensity on the first film. 7. If signal intensity was too high, wait up to 30 minutes before re-exposing. For detailed protocol, please Email us at [email protected] to request the detailed protocol. This product is for laboratory research ONLY and not for diagnostic use 2006 SignaGen Laboratories References: (1) Thorpe, G.H.G. and Kricka, L.J., Me thods in Enzymology, 133:331-353 (1986) (2) Thorpe, G.H.G., Kricka, L.J., Mosel ey, S.B. and Whitehead, T.P., Clin. Chem., 31(8):1335-1341 (1985) (3) Riko, I., et al, Analytical Biochemistry, 231:170-174 (1995) Storage: Upon arrival store this produ ct at 4 °C. Product shipped at cold-pa ck filled foam box.

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LumiGOLD™ ECL Western Blotting Detection Kit Standard Pack 250 ml Large Pack 1000 ml. 15875 Gaither Drive Gaithersburg, MD 20877 FAX. 301-560-4919 TEL. 301-330-5966 Toll Free. 1-866-918-6812 Email: [email protected] Web: www.signagenlabs.com. - PowerPoint PPT Presentation

TRANSCRIPT

Cat

# S

309

Sto

re a

t 4

0C LumiGOLD™ ECL Western Blotti

ng Detection Kit

Standard Pack 250 ml

Large Pack 1000 ml

15875 Gaither Drive

Gaithersburg, MD 20877

FAX. 301-560-4919

TEL. 301-330-5966

Toll Free. 1-866-918-6812

Email: [email protected]

Web: www.signagenlabs.com

Description:LumiGOLD™ ECL Kit is a complete kit with ready-to-use reagents for chemiluminescent detection of immobilized proteins (Western blotting) or immobilized nucleic acids (Southern or Northern), conjugated with HRP directly or indirectly.The use of enhanced chemiluminescence was introduced by Thorpe and Kricka (1,2). In the presence of hydrogen peroxide (H2O2 ), Horseradish peroxidase (HRP) catalyzes the oxidation of cyclic diacylhydrazides, such as luminol. Immediately following the oxidation, the luminol is inan excited state (intermediate reaction product), which decays to the ground state by emitting light. Strong enhancement of the light emission is produced by enhancers, such as phenolic compounds. Using this method, it is possible to detect membrane immobilized specific antigens, or sequences of nucleic acids, labeled directly with HRP or indirectly with HRP-labeled antibodies/streptavidin.

Quick Protocol:1. Mix an equal volume of LumiGOLD™ ECL Kit Reagent A and Reagent B to give sufficient solution to cover the membrane (0.1ml / cm2). Let the detection mix equilibrate for 2 minute at room temperature.2. Drain the excess buffer from the washed blots. Do not let the membrane dry out. Add the detection mix directly to the blot (protein side up). Incubate for 2 minute at roomtemperature. 3. Drain off excess detection mix and wrap the membrane in saran wrap. Gently remove air pockets.4. Place the blots immediately, protein side up, in the film cassette. Switch off the lights and use red safety light. Place a sheet of film on the blot, close the cassette and expose for 30-90 seconds or longer.5. Replace the exposed film with a new one, close the cassette and develop the first exposed film.6. Expose the second film for a suitable time according to the signal intensity on the first film.7. If signal intensity was too high, wait up to 30 minutes before re-exposing.

For detailed protocol, please Email us at [email protected] to request thedetailed protocol.

This product is for laboratory research ONLY and not for diagnostic use

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References:(1) Thorpe, G.H.G. and Kricka, L.J., Methods in Enzymology, 133:331-353 (1986)(2) Thorpe, G.H.G., Kricka, L.J., Moseley, S.B. and Whitehead, T.P., Clin. Chem.,31(8):1335-1341 (1985)(3) Riko, I., et al, Analytical Biochemistry, 231:170-174 (1995)

Storage: Upon arrival store this product at 4 °C. Product shipped at cold-pack filled foambox.

mailto:[email protected]

http://www.signagenlabs.com/

mailto:[email protected]

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