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Wählen Sie konfigurierbare Panels & Premixed-Kits - ODER - Kits für die zelluläre Signaltransduktion & MAPmates™
Konfigurieren Sie Ihre MILLIPLEX® MAP-Kits und lassen sich den Preis anzeigen.
Konfigurierbare Panels & Premixed-Kits
Unser breites Angebot enthält Multiplex-Panels, für die Sie die Analyten auswählen können, die am besten für Ihre Anwendung geeignet sind. Unter einem separaten Register können Sie das Premixed-Cytokin-Format oder ein Singleplex-Kit wählen.
Kits für die zelluläre Signaltransduktion & MAPmates™
Wählen Sie gebrauchsfertige Kits zur Erforschung gesamter Signalwege oder Prozesse. Oder konfigurieren Sie Ihre eigenen Kits mit Singleplex MAPmates™.
Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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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.
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Gewähltes Kit
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96-Well Plate
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Weitere Reagenzien hinzufügen (MAPmates erfordern die Verwendung eines Puffer- und Detektionskits)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Platzsparende Option Kunden, die mehrere Kits kaufen, können ihre Multiplex-Assaykomponenten in Kunststoffbeuteln anstelle von Packungen erhalten, um eine kompaktere Lagerung zu ermöglichen.
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7490-OP
Sigma-AldrichOmniPur® Silver Pro Stain Kit - Calbiochem
OmniPur® Silver Pro Stain Kit - Calbiochem Analysenzertifikate
Titel
Chargennummer
7490-OP
Datenblatt
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.
Revision
24-May-2011 JSW
Description
OmniPur® Silver Pro Stain Kit is an extremely sensitive colorimetric staining procedure for the detection of subnanogram amounts of proteins resolved on polyacrylamide gels. With high sensitivity and very low background, it is ideal for the visualization of protein bands in dilute samples or for the detection of proteins present in trace amounts. It is generally about 100-fold more sensitive than the commonly used Coomassie® Blue stain1.
The 4 hour staining procedure begins with a fixation step to reduce band diffusion and remove interfering substances. A sensitization process, which enhances binding of silver ions to proteins, follows fixation. The staining step allows silver ions to permeate the gel and bind preferentially to sulfhydryl and carboxyl side chains. In the final development step, silver ions are reduced to metallic silver to form precipitates that are visible as brown bands within the gel.
Staining intensity varies by the type of protein in the sample. Acidic proteins and proteins with highly negatively charged sulfated sugar residues such as proteoglycans and mucins are not readily detected by silver stains.
Recommended reaction conditions
Materials supplied: • 10X Sensitizer Solution, 1 bottle, 250 ml • 10X Silver Stain Solution, 1 bottle, 250 ml • 5X Developer Solution, 2 bottles, 250 ml each
Additional Materials Required but Not Supplied: • Methanol • Ethanol • 37% Formaldehyde • Glacial acetic acid • Distilled or deionized water
Precautions and Recommendations: • All incubations should be conducted with continuous agitation on a shaker. • Solutions should be freshly prepared on day of use. • All steps should be performed at room temperature. • Water must be deionized and distilled. • Staining should be carried out in clean glass containers with sufficient quantities of solution to immerse the gel and allow it to move freely during agitation. Use separate containers for each gel. • Powder-free gloves should be worn during the procedure. Do not touch gel plates, staining dishes or gels with bare hands as prints will be visible on the stained gels. Rinse gloves in water between each step. • If crystals form in the developer solution during storage, warm gently until they re-dissolve.
Reagent Preparation • Fixative (100 ml): prepare Fixative as follows: • 50 ml Methanol • 12 ml Glacial Acetic Acid • 1.35 ml 37% Formaldehyde • 36.65 ml Distilled/Deionized Water
• 35% Ethanol (300 ml): prepare 35% ethanol as follows: • 81 ml 95% Ethanol • 219 ml Distilled/Deionized Water
• 1X Sensitizer Solution (100 ml): prepare 1X Sensitizer Solution as follows: • 10 ml
10X Sensitizer Solution • 90 ml Distilled/Deionized Water
• 1X Stain Solution (100 ml): prepare 1X Stain Solution as follows: • 10 ml 10X Stain Solution • 90 ml Distilled/Deionized Water
• 1X Developer Solution (100 ml): prepare 1X Developer Solution as follows: • 20 ml 5X Developer Solution • 80 ml Distilled/Deionized Water
• Stop Solution (100 ml): prepare Stop Solution as follows: • 50 ml Methanol • 12 ml Glacial Acetic Acid • 38 ml Distilled/Deionized Water
Detailed Protocol 1. Fix gel in 100 ml of Fixative 2 hr to overnight. 2. Wash 3 times, 20 min per wash, in 35% Ethanol. 3. Incubate gel in 100 ml 1X Sensitizer Solution for 2 min. 4. Wash 3 times, 5 min per wash, in distilled/deionized water. 5. Incubate gel for 20 min in 1X Stain Solution. 6. Wash gel 2 times, 1 min per wash, in distilled/deionized water. 7. Incubate gel in 1X Developer Solution until bands become visible (approximately 10 min for full development). Bands should appear dark brown against a pale background.
Note: • The rate of band development is temperature dependent. • If the gel is over-developed, artifacts are present, the background is too dark, or if the bands are over-stained, the gel can be de-stained and restained as desired.
8. Incubate for 20 min in Stop Solution to prevent further color development. 9. Store at 4°C in 1% Acetic Acid. 10. Gels may be photographed on a bright white light box.
Troubleshooting Guide
A.Background is too dark: 1. Residual acid in gel • Increase washing time after the fixation step.
2. Poor quality acrylamide • Acrylamide quality can affect the background appearance of a silver-stained gel. Use ultrapure grade acrylamide.
3. Poor quality water • Use deionized/distilled water in all solutions.
B. Negative staining 1. Excess protein in bands. • Reduce the amount of protein applied to gel.
C. Streaking or yellow background 1.Excess reducing agents such as 2-mercaptoethanol or DTT. • Reduce the amount of reducing agent in sample buffer.
D. Artificial bands with apparent molecular weights between 50-70 kDa 1. Excess amounts of reducing agents such as 2-mercaptoethanol or DTT. • Lower the amount of reducing agent in sample buffer.
