The Product Information Sheet recommends a protease inhibitor suitable for mammalian cell and tissue extracts, such as product numbers P8340, PIC0002, or I3786, as listed under "Reagents and Equipment Required but Not Provided." After adding the CelLytic M reagent, a 15-minute incubation on a shaker is recommended. The lysate should not be vortexed.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/113/033/c2978bul-ms.pdf
C2978
CelLytic™ M
Cell Lysis Reagent, Suitable for Mammalian cell lysis and protein solubilization.
Synonym(e):
Whole-cell protein extraction solution
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NACRES:
NA.56
UNSPSC Code:
41116134
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Dieser Artikel | |||
|---|---|---|---|
| form solution | form powder | form - | form - |
| storage temp. room temp | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
General description
CelLytic M ist eine firmeneigene Detergenslösung zur effizienten Ganzzell-Proteinextraktion aus kultivierten Säugerzellen. Sie ermöglicht eine effiziente und schnelle Zelllyse und Solubilisierung von Proteinen sowohl für suspendierte als auch für adhärente Zellen. Lysate können in vielen nachgelagerten Anwendungen verwendet werden, ohne dass CelLytic M entfernt werden muss, z. B. in Reportergen-Assays, Western-Blot-/Immunpräzipitations-Assays, Electrophoretic Mobility Shift Assays (EMSA), Phosphatase-Assays und Kinase-Assays.
Features and Benefits
- Effizient: Um bis zu 50 % effizienter als Einfrier-/Auftau-, Ultraschall- und andere Produkte
- Nicht denaturierend: Behindert keine nachgelagerten Anwendungen wie die Immunpräzipitation, Kinase- und Phosphatase-Assays, Reportergen-Assays und Gel-Shift-Assays.
- Praktisch: Gebrauchsfertiges Reagenz erfordert kein Abschaben von Kulturplatten
- Schnell: Schnelle Zelllyse bei Raumtemperatur.
Preparation Note
Die Effizienz des Reagenz CelLytic M für die Proteinextraktion wurde u. a. an HeLa, CHO, COS, HL-60, Jurkat, A431, PC-12 und Endothelzellen aus der Rinderaorta (BAEC) getestet.
Analysis Note
125 μL CelLytic M für 106-107 suspendierter Zellen verwenden. Für adhärente Zellen 500–1.000 μL für eine 100-mm-Platte bzw. 200–400 μL für eine 35-mm-Platte verwenden.
Legal Information
CelLytic is a trademark of Sigma-Aldrich Co. LLC
Lagerklasse
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| C2978-250ML | 04061838445568 |
| C2978-40ML | 04061838248190 |
| C2978-10ML | 04061838248183 |
| C2978-50ML | 04061838445575 |
| C2978-60ML | 04061838112828 |
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1. Do you recommend supplementing CelLytic M lysis buffer with a protease inhibitor cocktail? 2. Is just chemical lysis typically sufficient in this buffer or you suggest any physical method in addition such as vortexing?
1 answer-
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Does CelLytic™ M has EDTA? Is it compatible with Phos-Tag gel?
1 answer-
The formulation does not contain EDTA or chelators. There is no known reason for it to be incompatible with Phos-Tag gel, but it has not been tested for this specific application.
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Does the CelLytic M (Cat # 2978) contain DTT, and if it does, at what concentration?
1 answer-
As mentioned in the 'DESCRIPTION' section, the composition of this product is considered proprietary and may not be shared. This product does not contain any denaturants except for a proprietary mild detergent. Lysates obtained with this product may be immediately run on a SDS PAGE after the addition of loading buffer.
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Is the product compatible with the BCA assay? Additionally, are there any denaturants in the formulation?
1 answer-
CelLytic™ M is suitable for Bradford Assay and BCA assays. For the Lowry method, there is a background issue that is problematic for low protein concentrations, but may be subtracted with high protein concentrations.
CelLytic™ M does not contain any primary amines; it is a buffered solution (pH 7.6) which has no denaturants except for a mild detergent. The exact formulation is proprietary. This product may be diluted or dialyzed if it is found to have interference in your particular assay.Helpful?
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What is the critical micelle concentration (CMC) of the product?
1 answer-
Unfortunately, the CMC value has not been tested for this product.
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Does the CelLytic M (Cat. No.: 2978) contain SDS, and if it does, at what concentration?
1 answer-
The product with the code C2978, CelLytic M, does not contain SDS. Instead, it contains a mild detergent in a buffer.
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Is the C2978 lysis buffer suitable for viral DNA extraction from cultured mammalian cells (fewer than 25,000 cells/ul) and subsequent qPCR?
1 answer-
The cellular DNA is centrifuged down and will not be present in the supernatant along with the proteins. To isolate the DNA, a different kit is necessary.
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Has CelLytic been tested for neutralizing HIV or use in a BSL3 facility?
1 answer-
We have not tested CelLytic for neutralizing HIV nor used in a BSL3 facility.
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Will CelLytic™ M, Product No. C2978, interfere with protein concentration assays such as Bradford, Lowry and BCA?
1 answer-
We have tested and found CelLytic™ M to be compatible for Bradford Assay and BCA assays. For the Lowry method, there is a background issue that is problematic for low protein concentrations, but may be subtracted with high protein concentrations.
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Is there any denaturants in Product C2978, CelLytic™ M that can interfere with my downstream applications?
1 answer-
CelLytic™ M does not contain any primary amines; it is a buffered solution (pH 7.6) which has no denaturants except for a mild detergent. The exact formulation is proprietary. This product may be diluted or dialyzed if it is found to have interference in your particular assay.
Helpful?
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