Product Class: Other

RecAf
NEBU recombinant 37 65 Heat

This product has been replaced with RecA (NEB #M0249).

RecAf is His-tagged. Native RecA is also available as a separate product.

  • Replaced by RecA on December 15, 2021
  • Catalog # M0355 was discontinued on December 15, 2021

Product Introduction

RecAf is a N-terminal 6X His tagged recombinant protein.  E. coli RecA is necessary for genetic recombination, reactions involving DNA repair and UV-induced mutagenesis. RecA promotes the autodigestion of the lexA repressor, umuD protein and lambda repressor. Cleavage of LexA derepresses more than 20 genes. In vitro studies indicate that in the presence of ATP, RecA promotes the strand exchange of single-strand DNA fragments with homologous duplex DNA. The reaction has three distinct steps: (i) RecA polymerizes on the single-strand DNA, (ii) the nucleo­protein filament binds the duplex DNA and searches for a homologous region, (iii) the strands are exchanged.
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Catalog # Size Concentration

Product Information

Description
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0355S     -20    
      RecAf M0355SVIAL -20 1 x 0.1 ml 2 mg/ml
      Rec A Reaction Buffer B0355SVIAL -20 1 x 1 ml 10 X
  • M0355L     -20    
      RecAf M0355LVIAL -20 1 x 0.5 ml 2 mg/ml
      Rec A Reaction Buffer B0355SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Reaction Conditions

1X Rec A Reaction Buffer
Incubate at 37°C

1X Rec A Reaction Buffer
70 mM Tris-HCl
10 mM MgCl2
5 mM DTT
(pH 7.6 @ 25°C)

Heat Inactivation

65°C for 20 minutes

Application Features

  • Visualization of DNA structures with electron microscopy (3)
  • D-loop mutagenesis (4)
  • Screening libraries using RecA-coated probes (5,6)
  • Cleavage of DNA at any single predetermined site (7,8,9)
  • RecA mediated affinity capture for full length cDNA cloning (10,11)

Product Notes

  1. ATPγS which is required for triple helix formation is not supplied.
  2. Heat Inactivation: 65°C for 20 min

References

  1. West, S.C. (1992). Ann. Rev. Biochem. 61, 603-640.
  2. Zhumabayeva, B. et al. (1990). Biotechniques. 27, 834-845.
  3. Zhumabayeva, B. et al. (2001). Biotechniques. 30, 512-520.
  4. Pace, C.N. et al. (1995). Protein Sci. 4, 2411-2423.
  5. Radding, C.M. (1991). J. Biol. Chem. 266,
  6. Wasserman, S.A. and Cozzarelli, N.R. (1985). Proc. Natl. Acad. Sci. USA. 82, 1079-1083.
  7. Shortle, D. et al. (1980). Proc. Natl. Acad. Sci. USA. 77, 5375-5379.
  8. Honigberg, S.M. et al. (1986). Proc. Natl. Acad. Sci. USA. 83, 9586-9590.
  9. Rigas, B. et al. (1986). Proc. Natl. Acad. Sci. USA. 83, 9591-9595.
  10. Ferrin, L.J. and Camerini-Otero, R.D. (1991). Science. 254, 1494-1497.
  11. Koob, M. et al. (1992). Nucleic Acids Res. 20, 5831-5836.
  12. Koob, M. et al. (1992). In R. Wu(Ed.), Methods in Enzymology. 216, 321-329. San Diego: Academic Press.

Protocols, Manuals & Usage

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. What buffer should be used with RecAf Protein?
  2. Is magnesium required for triple strand formation using RecAf Protein?
  3. Is ATP gamma-S required for triple strand formation using RecAf Protein?
  4. Can the RecAf Protein be heat inactivated?
  5. Is the structure of the target DNA important when using RecAf Protein?
  6. How much RecAf Protein should be added to the single stranded DNA to coat it?
  7. What length single stranded oligo should be used with RecAf Protein?

Tech Tips

It’s N-term 6XHis tagged RecA with moderately stronger ssDNA binding ability compared to wt RecA. A possible reason might be the increased pI due to the attachment of the 6XHis-tag.

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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