secondary antibody

Enhanced Chemiluminescent (ECL) Detection Kit (Mouse IgM)

EK1005

1 kit, with removable strips.

Enabled

Enhanced Chemiluminescent (ECL) Detection Kit (Mouse IgM)

Secondary Antibodies, Western Blotting Reagents, All-In-One Western Blot Kits

No

Enhanced Chemiluminescent Reagent Kit (Mouse IgM) for Western Blotting, with the specific secondary antibody and all the reagents needed for performing WB. 96wells/kit, with removable strips.

Enhanced Chemiluminescent Reagent Kit (Mouse IgM). 96wells/kit, with removable strips.

Overview Product Name Enhanced Chemiluminescent Reagent Kit (Anti-Mouse IgM) Synonyms ECL Anti-Mouse Antibody Detection Reagent System; ECL Western Blotting Substrate Reagent Kit (Anti-Mouse IgM); ECL-WB Substrate Reagent System (Anti-Mouse IgM); Enhanced Chemiluminescent Western Blot Related HRP-Conjugated Goat Anti-Mouse IgM Assay System Description Enhanced Chemiluminescent Reagent Kit (Anti-Mouse IgM) is a four-component system for sensitive detection of membrane-immobilized proteins on western blots probed with mouse primary antibodies using HRP-conjugated anti-mouse secondary antibodiesof class M and enhanced chemiluminescent peroxidase substrate reagents. Application Western blot Pack Size 1 kit (for 1000 cm² membrane) Assays Per Kit Sufficient for covering up to 1000 cm² of PVDF or nitrocellulose membrane Reagent Type Protein assay kit; ECL Western Blotting related Kit Components* Kit Component Component Description Size Blocking buffer Nonfat dry milk protein powder 10 g HRP-conjugated secondary antibody, affinity purified, solid phase adsorbed HRP-conjugated goat anti-mouse IgM 0.1ml Enhanced chemiluminescent (ECL) reagent A HRP chemiluminescent substrate: luminol-based; enhancer 5 ml Enhanced chemiluminescent (ECL) reagent B chemiluminescent substrate oxidizing agent 5 ml . *Additional components can be purchased. If you need extra of the above components please order them together to avoid additional shipping charges. Assay Information Sample Type SDS-PAGE separated-, membrane-immobilized-, primary-antibody-probed proteins from cell/tissue lysates Assay Type Immunoassay Assay Purpose Detect target protein; Quantify target protein level Technique Immunodetection of target protein by HRP-conjugated secondary antibody Detection Method Chemiluminescent Emission 428 nm, blue glow Equipment Needed WB instrumentation; X-ray film cassette or a charge-coupled device (CCD) imager Assay Sensitivity Femtomole and picogram protein quantities Properties Form Supplied Blocking buffer: Dry powder . HRP-Antibody conjugate: Concentrated stock solution, 0.1 mg/ml, in 0.01M PBS (pH7.4) and 50% glycerol . ECL reagents A and B: Concentrated stock solutions, 20x Recommended Working Concentration See protocol in Datasheet Storage 4˚C for one year Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE Main Advantages Specific High signal-to-noise ratio Sensitive Up to 10 times more sensitive than colorimetric methods Quantitative Linear response in the range 0.2 to 2 OD units Stable The light output signal remains detectable up to several hours after incubation Reliable Possibility for repeated exposure to X-ray film until obtain optimal results Storable Immediate hard copy data - avoid fading of results Reuseable Allowing reprobing on membranes up to 10 times - saves time and materials Fast Short reaction times; few sec to few min of exposure enough to detect signal Non-radioactive and Non-toxic No specialized facilities needed Background . Chemiluminescence refers to the light emitted by a chemical reaction and chemiluminescent systems have emerged as the best all-around technique for the revelation step of blotting-based analyses. Horseradish Peroxidase (HRP) Conjugated Secondary Antibodies are the most popular antibody conjugates and are widely used to obtain chemiluminescent results in western and other immunoblotting, ELISA, and immunochemistry. In the chemiluminescence reaction Horseradish peroxidase (HRP) catalyzes the oxidation of cyclic diacylhydrazides, such as luminol, or acridine derivatives in the presence of hydrogen peroxide (H2O2) acting as activator or oxidizing agent. Immediately following the oxidation, the luminol is in an excited state (intermediate reaction product), which decays to the ground state by emitting light. Strong up to 1000-fold enhancement of the light emission is produced by enhancers, such as phenolic compounds, making the light emission much prolonged and very intense. This allows detection of protein at a level of 1-10 pg. Enhanced Chemiluminescent (ECL) Reagent Kit is an immunodetection system that utilizes horseradish peroxidase (HRP) conjugated anti-mouse, anti-rabbit, anti-goat, or anti-rat secondary antibodies for ECL western blot detection of proteins. Proteins from various samples such as cell lysates are first separated within an acrylamide gel under adapted electric current (electrophoresis) and then transferred onto a PVDF or nitrocellulose membrane (transfer). The membrane containing the proteins is then incubated with specific primary antibodies which bind to their specific target(s) (capture) and subsequently with Horseradish Peroxidase (HRP)-conjugated secondary antibodies which bind to the primary antibodies (detection). Addition of chemiluminescent reagents (HRP luminol-based substrate, enhancer and activator) results in a chemiluminescent signal that can be captured using photographic or other imaging methods (revelation). The peroxidase-catalyzed oxidation of luminol produces a glow-type light emission at 428 nm. The intensity of light output is proportional to peroxidase activity and is a measure of the number of enzyme molecules reacting and thus of the amount of target antigen.

1 kit, with removable strips.

No

4 C for one year.

1. Blocking buffer: 10g protein dry powder (nonfat dry milk). 2. HRP-conjugated goat anti-mouse IgM: Affinity purified antibody. 3. Enhanced chemiluminescent chromogenic reagent, containing: -Chromogenic reagent A: 5ml. 20x. -Chromogenic reagent B: 5ml. 20x.

WB

Enhanced Chemiluminescent Reagent Kit (Anti-Mouse IgM) components: . Blocking buffer: 10g . HRP-conjugated goat anti-mouse IgM: 0.1 ml . Chemiluminescent reagent A: 5 ml . Chemiluminescent reagent B: 5ml Enhanced Chemiluminescent (ECL) Detection Mechanism

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