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B7310

Sigma-Aldrich

CelLytic ™ B 细胞裂解试剂

For bacterial cell lysis, 2× concentrate

同義詞:

bacterial cell lysis reagent

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About This Item

MDL號碼:
MFCD02097909
分類程式碼代碼:
41116134
NACRES:
NA.56
暫時無法取得訂價和供貨情況

應用

两性离子洗涤剂的专利、非变性配方，用于细菌细胞裂解和重组蛋白提取。

特點和優勢

  • 比传统方法如超声法和溶菌酶法提取蛋白质效率高
  • 可放大 1-25 克细菌细胞浆
  • 不干扰下游应用，如亲和层析、IP 和免疫印迹
  • 与蛋白酶抑制剂、抑制剂混合物、离液剂、盐、螯合剂、还原剂相容

其他說明

2× concentrate of a detergent blend formulated in 40 mM Trizma® HCl (pH 8.0).

法律資訊

此产品受美国专利号7,282,475 B2管辖，仅针对研究用途出售。如需商业化使用，需另行征得许可。
CelLytic is a trademark of Sigma-Aldrich Co. LLC
Trizma is a registered trademark of Merck KGaA, Darmstadt, Germany

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類（WGK）

WGK 2

閃點（°F）

Not applicable

閃點（°C）

Not applicable

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分析證明 (COA)

Lot/Batch Number
0000392900
0000345849
0000345857
0000345849
0000345857

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Russel M Vincent et al.
ACS synthetic biology, 8(4), 675-685 (2019-03-12)
Using engineered initiator tRNA for precise control of protein translation within cells has great promise within future orthogonal translation systems to decouple housekeeping protein metabolism from that of engineered genetic systems. Previously, E. coli strain C321.ΔA. exp lacking all UAG stop

Questions

1–10 of 10 Questions  

  1. What is the difference between CelLytic B (Product Nos. B7435, B7310, C8740) and CelLytic B Plus  (Product Nos. CB0050 and CB0500)?

    1 answer

    1. CelLytic B Plus contains CelLytic B, Product No. B7435 as well as lysozyme, benzonase, and a protease inhibitor cocktail for a complete reagent for cell lysates for protein isolation.

      Helpful?

  2. Can I use the CelLytic™ B cell lysate in an activity assay?

    1 answer

    1. CelLytic B contains a zwitterionic detergent so the protein is active and can be used in protein activity assays.

      Helpful?

  3. Can I use CelLytic B for eukaryotic cells?

    1 answer

    1. CelLytic B was optimized for bacterial cells.  It may lyse some mammalian cells. CelLytic B 10X is a better choice for the lysis of yeast and mammalian cells due to the increased detergent concentration.

      Helpful?

  4. Can I use CelLytic B with the maltose binding protein system (MBP System)?

    1 answer

    1. CelLytic B cannot be used with the MBP system.  We have used this in the maltose binding protein system, and know that it will NOT work in this system, since any and all detergents will destroy binding of protein to the resin.

      Helpful?

  5. Can I concentrate my sample that was made using CelLytic B?

    1 answer

    1. If concentrating a sample, be aware that concentration using filter concentrators may increase the detergent concentration significantly and cause issues in downstream applications.  heoretically, ways to get around this are TCA precipitation, or diluting the extract below the CMC prior to concentration. This would be at least a 1/5 dilution, based on the following reference: Saito, S., and Tsuchiya, T., Characteristics of n-octyl beta-D-thioglucopyranoside, a new non-ionic detergent useful for membrane biochemistry. Biochem. J., 222(3), 829-832, 1984.

      Helpful?

  6. When using the CelLytic™ B lysis reagent, do I need to make a dilution prior to measuring protein by Bradford assay?

    1 answer

    1. When using the CelLytic™ B lysis reagent (B7435, B7310, C8740, and contained in kits CB0050 and CB0500), it is best to use a 1:10 dilution in order to reduce the risk of interference with the Bradford Reagent.  A 1:20 dilution would be even better, as long as the protein level remains within the standard curve.

      Helpful?

  7. What is the difference between CelLytic B (Product No. B7435), CelLytic B 2x (Product No. B7310) and CelLytic B 10x (Product No. C8740)?

    1 answer

    1. CelLytic B 2X has double the strength of CelLytic B for small volume extractions. This allows CelLytic B 2X to be used when a higher protein concentration or lower volume is required. The original formula, CelLytic B, requires 10-20 ml of the reagent for 1 gram of wet cell paste.  Only 5 ml of the CelLytic B 2X reagent is required to lyse and extract protein from 1 gram of wet cell paste.  CelLytic B 10X is provided as a concentrated mixture of the proprietary detergents found in CelLytic B, without any buffering component. This allows the user to customize the extraction reagent by choosing the detergent concentration and buffering components ideal for the extraction and purification of the protein(s) of interest. CelLytic B 10X is also suitable for the lysis of yeast and mammalian cells.

      Helpful?

  8. What bacterial cells can be used with CelLytic B?

    1 answer

    1. CelLytic B Reagent by itself (no lysozyme or benzonase) was optimized for E. coli strain BL21.  It also works with other common E. coli strains such as DH5?, JM109, and HB 101. The CelLytic B Reagent may be used on other similar bacterial cells. The CelLytic B Plus Kit is compatible with many more difficult to lyse Gram positive bacteria such as Bacillus and Streptococcus.

      Helpful?

  9. Can I obtain protein from inclusion bodies with CelLytic B?

    1 answer

    1. If the protein is found in inclusion bodies, you need to use an additional reagent, Product No. C5236, CelLytic IB to solubilize the proteins from the inclusion bodies.  CelLytic B is used to isolate the inclusion bodies from the bacteria.  CelLytic IB, Product No. C5236 can then be used to solubilize the proteins from the inclusion bodies.

      Helpful?

  10. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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