2A Peptide Antibody (3H4R) - BSA Free

Novus Biologicals | Catalog # NBP3-09171

Recombinant Monoclonal Antibody
Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Non-species specific

Cited:

Human, Mouse

Applications

Validated:

Western Blot

Cited:

Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 3H4R

Format

BSA Free
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Product Specifications

Immunogen

This Recombinant 2A Peptide Antibody (3H4) was developed against KLH-coupled synthetic peptide corresponding to 2A peptide of Thosea asigna virus (CGDVEENPG)

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for 2A Peptide Antibody (3H4R) - BSA Free

Western Blot Detection of 2A Peptide in HeLa Cells

Western Blot Detection of 2A Peptide in HeLa Cells

Total protein from SpCas9-T2A peptide transfected HeLa cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/ml anti-T2A in 5% non-fat milk in TBST and detected with an anti-mouse HRP secondary antibody using chemiluminescence.

Applications for 2A Peptide Antibody (3H4R) - BSA Free

Application
Recommended Usage

Western Blot

1 - 2 ug/ml

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: 2A Peptide

Initially discovered from the foot-and-mouth disease virus (FMDV, a picornavirus), 2A peptide is a self-cleaving peptide 18-22 amino acids long with a conserved C-terminal motif Asp-Val/Ile-Glu-X-Asn-Pro-Gly-Pro. All picornaviruses have 2A peptides but they differ in size and function. The FMDV genome is a single open reading frame encoding a polyprotein and 2A denotes a specific cleavage region within the polyprotein. Co-translational cleavage of the FMDV polyprotein is not mediated by a proteolytic mechanism but rather by "ribosomal skipping" or "StopGO" at the 2A/2B junction. When synthesis terminates at a sense codon, the polypeptide is released before resuming translation at the next codon (1, 2).

Developed for genetic engineering, 2A cleavage has been shown to function in a wide range of eukaryotic cells, and in vivo (3). The 2A peptide is inserted between the coding sequences of at least two genes, enabling the simultaneous expression of multiple proteins from a single plasmid construct. Four variants of 2A peptides have been used in gene expression systems including FMDV 2A (F2A), equine rhinitis A virus 2A (E2A), porcine teschovirus-1 2A (P2A) or Thoseaasigna virus 2A (T2A). Compared to other multi-gene co-expression systems such as IRES, 2A peptides results in stoichiometric expression of different proteins. However, cleaving efficiency and protein expression using the different 2A peptides varies, requiring optimization of polycistronic constructs for experiments.

References

1. Liu Z, Chen O, Wall J, Zheng M, Zhou Y, Wang L, Vaseghi H, Qian L, Liu J. (2017). Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. Scientific Reports. 7. 10.1038/s41598-017-02460-2.

2. Luke GA, Escuin H, Felipe PD, Ryan MD. (2009) 2A to the Fore - Research, Technology and Applications, Biotechnology and Genetic Engineering Reviews. 26(1): 223-260.

3. Markus BM, Bell GW, Lorenzi HA, Lourido S. (2019) Optimizing Systems for Cas9 Expression in Toxoplasma gondii. mSphere. 4(3): e00386-19.

Alternate Names

2A cleavage peptide, 2A peptide, 2A Peptide Antibody, 2A peptide tag, 2A sequence, 2A-like peptide, antibody for 2A-containing recombinant proteins, FMDV 2A-peptide, FMDV 2A-peptide antibody, MDV 2A-peptide, P2A peptide, P2A tagged proteins, peptide 2A sequences, self-cleaving 2A peptide, T2A peptide, T2A tagged proteins, Thosea asigna virus 2A, Thosea asigna virus 2A antibody

Additional 2A Peptide Products

Product Documents for 2A Peptide Antibody (3H4R) - BSA Free

Certificate of Analysis

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Product Specific Notices for 2A Peptide Antibody (3H4R) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for 2A Peptide Antibody (3H4R) - BSA Free

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