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.
Sinónimos:
Whole-cell protein extraction solution
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UNSPSC Code:
41116134
NACRES:
NA.56
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Este artículo | C1990 | CE0050 | CB0500 |
|---|---|---|---|
| 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 is a proprietary detergent solution designed for efficient whole-cell protein extraction from cultured mammalian cells. It enables efficient and rapid cell lysis and solubilization of proteins for both suspension and adherent cells. Lysates can be used in many downstream applications without removing the CelLytic M such as reporter gene assays, Western blots/immunoprecipitation, electrophoretic mobility shift assays, phosphatase assays and kinase asssays.
Features and Benefits
- Efficient: Up to 50% more efficient than freeze thaw, sonication and other products
- Non-denaturing: Does not interfere in downstream applications such immunoprecipitation, kinase and phosphatase assays, reporter gene assays and gel shift assays
- Convenient: Ready-to-use reagent requires no scraping from culture plates
- Fast: Rapid cell lysis at room temperature.
Preparation Note
CelLytic M reagent efficiency for protein extraction has been tested on, but not limited to, HeLa, CHO, COS, HL-60, Jurkat, A431, PC-12, and Bovine Aorta Endothelial Cells (BAEC).
Analysis Note
Use 125 μl CelLytic M for 106-107 of suspended cells. For adherent cells, use 500-1,000 μL for a 100 mm plate; 200-400 μL for a 35 mm plate.
Legal Information
CelLytic is a trademark of Sigma-Aldrich Co. LLC
Clase de almacenamiento
12 - Non Combustible Liquids
wgk_germany
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
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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.
Helpful?
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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.
Helpful?
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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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