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Key Product Details

Species Reactivity

Non-species specific

Applications

Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 3H4

Concentration

1.0 mg/ml

Product Summary for 2A Peptide Antibody (3H4)

Immunogen

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

Specificity

This 2A Peptide Antibody (3H4) peptide antibody is specific for T2A and P2A tagged proteins. Specificity to F2A and E2A proteins has not been tested.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for 2A Peptide Antibody (3H4)

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.
Detection of 2A Peptide in NIH-3T3 Cells

Detection of 2A Peptide in NIH-3T3 Cells

2A Peptide Antibody used for Western Blotting. NIH-3T3 cells were stably integrated with a lentiviral plasmid expressing a fluorescent protein-P2A protein. Two different viral titers were used in this experiment. Image from verified customer review.

Applications for 2A Peptide Antibody (3H4)

Application
Recommended Usage

Western Blot

1 ug/ml
Application Notes
Use in flow cytometry reported in PMID: 36356599 and customer review. Novus does not validate this product in flow cytometry and is unable to guarantee its performance.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP2-59627 in the following applications:

Published Applications

Read 15 publications using NBP2-59627 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

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

Product Documents for 2A Peptide Antibody (3H4)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for 2A Peptide Antibody (3H4)

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.

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FAQs for 2A Peptide Antibody (3H4)

Showing  1 - 2 of 2 FAQs Showing All
  • Q: Are there any cleavage sites on 2A peptide, Clone 3H4?

    A: Scientific articles have referenced the recognition of 2A sequence derived from foot and mouth piconavirus (VKQTLNFDLLKLAGDVESNPG*P) with cleavage site between G and P.

  • Q: How long are the 2A sequences and does it encode more than one protein?

    A: The sequence is approximately 20 amino acids long based on the source of virus. 2A peptides allow several proteins to be encoded as polyproteins, which then separate into component proteins upon translation.

Showing  1 - 2 of 2 FAQs Showing All
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