Optimal dilution of this antibody should be experimentally determined. 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.
Reviewed Applications
Read 1 Review rated 5 using NBP2-59627AF647 in the following applications:
Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.
Alternate Names for 2A Peptide Antibody (3H4) [Alexa Fluor® 647]
2A cleavage peptide
2A Peptide Antibody
2A peptide tag
2A peptide
2A sequence
2A-like peptide
antibody for 2A-containing recombinant proteins
FMDV 2A-peptide antibody
FMDV 2A-peptide
MDV 2A-peptide
P2A peptide
P2A tagged proteins
peptide 2A sequences
self-cleaving 2A peptide
T2A peptide
T2A tagged proteins
Thosea asigna virus 2A antibody
Thosea asigna virus 2A
Background
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.
Limitations
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.
Publications for 2A Peptide Antibody (NBP2-59627AF647) (0)
There are no publications for 2A Peptide Antibody (NBP2-59627AF647).
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Review for 2A Peptide Antibody (NBP2-59627AF647) (1)
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Cells were permeabilized with 4% PFA for 15 mins at room temperature and stained with NBP2-59627AF647 for 30 mins at room temperature. ***Bio-Techne Response: This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.***
Product General Protocols
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FAQs for 2A Peptide Antibody (NBP2-59627AF647). (Showing 1 - 2 of 2 FAQ).
Are there any cleavage sites on 2A peptide, Clone 3H4?
Scientific articles have referenced the recognition of 2A sequence derived from foot and mouth piconavirus (VKQTLNFDLLKLAGDVESNPG*P) with cleavage site between G and P.
How long are the 2A sequences and does it encode more than one protein?
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.
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