Cusabio N-terminal Myc-tagged Recombinant

przez | 7 lutego, 2022

What is a Myc tag?

A Myc tag is a polypeptide protein tag derived from the product of the c-Myc gene that can be fused to the C- or N-terminus of a protein through recombinant DNA technology. It can facilitate the affinity purification of the tagged protein, as well as the isolation and detection of the tagged protein.

What is the amino acid sequence of the Myc-tag peptide?

The peptide sequence of the myc tag is EQKLISEEDL (or -Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-).

What is the molecular weight of the Myc tag?

The Myc-tag contains 10 amino acids, and the molecular weight of the Myc-tag peptide (EQKLISEEDL-tag) is 1202 Da.

Why use a Myc tag for protein expression and production?

Adding a Myc tag to the C or N terminus of a protein can facilitate its detection, isolation, and purification by Myc tag-specific antibodies. This is particularly useful for proteins without a detection antibody or with an unknown cellular location. Sino Biological has developed Myc-tag specific antibodies for ELISA, Western Blotting, immunofluorescence and immunoprecipitation for the detection of Myc-tagged protein. It can also be conjugated to agarose beads for affinity purification of Myc-tagged proteins.

How are Myc-tagged proteins purified?

The N-terminal Myc-tagged Recombinant protein can be affinity purified directly from cell culture lysate or supernatant. Myc-tagged protein is bound to conjugated Myc-tag-specific monoclonal antibody on an agarose gel. After washing away residual impurities, bound Myc-tag proteins can be eluted from the affinity column by a high concentration of Myc-tag peptide or by a low pH buffer.

How is it separated from Myc-tag after purification?

In some applications, it is desirable to remove the Myc tag, eg for protein crystallization. To allow cleavage of the Myc tag, it is necessary to design a protease cleavage site between the tag and the protein. An EK cleavage site behind the Myc tag (Myc-EK site protein structure) may allow complete removal of the Myc tag and cleavage site, leaving no additional amino acids after specific cleavage of the Myc tag. For more information on cleavage site and tag removal by EK and HRV-3C protease, see Enterokinase (EK), HRV-3C (Human Rhinovirus Protease).

Myc-tag protein production and purification service

Complete services: From gene cloning/synthesis to purified proteins

Host cells

Choices of expression hosts:

  • Transient transfection in HEK293 cells
  • Transient transfection in CHO cells
  • Stable expression in CHO cells
  • Yeast cell expression system
  • Baculovirus/insect cell expression system
  • E. coli. microbial expression system

Myc tag design: Myc tag (EQKLISEEDL tag) at N- or C-terminus

Cleavage site: Option to design a cleavage site between the tag and the protein

Protease Cleavage Enzyme Selection: Enterokinase (EK), HRV-3C (Human Rhinovirus Protease), TEV

Purification: Myc-tag (EQKLISEEDL-tag) specific antibody conjugate affinity group Myc-tag Affinity purification resin

Deliverable: Purified recombinant proteins (amounts to be specified by the client)

Quality specification: typically >90% purity per SDS page

Typical schedule: 8-12 weeks

Myc-tag (EQKLISEEDL-tag) has been widely used in the expression and production of recombinant proteins. Myc-tag allows simple and efficient affinity purification of tagged protein using Myc-tag specific antibodies conjugated to agarose beads. The Myc tag (EQKLISEEDL tag) can also be used for detection in Western Blot by using a Myc tag-specific antibody. Myc-tag can be added to the C- or N-terminus of a protein for expression in virtually all expression systems.

Sino Biological has successfully produced many Myc-tagged proteins and purified them with Myc-tagged affinity chromatography. We have a lot of experience in designing Myc-tag according to protein structure and handling any issues that arise from the protein purification process. Sino Biological has also developed highly sensitive Myc-tag (EQKLISEEDL-tag) specific antibodies for detection in western blot, ELISA assay, immunofluorescence and immunoprecipitation. We have also developed our own Myc-tag affinity purification resin (EQKLISEEDL-tag). For more information on our Myc-tag epitope antibodies (EQKLISEEDL-tag), see Tag Antibodies. Myc-tag antibodies can be purchased directly from our website.

Our track record in recent years in successfully producing around 6,000 bulk recombinant proteins and over 12,000 antibodies has attracted top 10 multinational pharmaceutical customers as well as small biotech companies worldwide through word of mouth. mouth, since we haven’t. done a lot of business development or advertising in the past. Also, we are starting to promote our protein production services to all research institutions and companies around the world, as we believe that more customers can greatly benefit from our ability and experience.