2A Peptide Antibody (3H4) - BSA Free

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.

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.

• Reactivity: All-NA
• Applications: WB, ICC/IF, IP
• Clone: 3H4
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

2A Peptide Antibody (3H4) - BSA Free Summary

• Additional Information: Non-recombinant Monoclonal Antibody
• 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.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunoprecipitation
  • 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for 2A Peptide Antibody (3H4) - BSA Free

• 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 (NBP2-59627)(17)

Publications using NBP2-59627

• Snyder AJ, Agbemabiese CA, Patton JT. et Al. Production of OSU G5P[7] Porcine Rotavirus Expressing a Fluorescent Reporter via Reverse Genetics Viruses 2024-03-07 [PMID: 38543776]
• Zhang H, Papadaki S, Sun X et al. Quantitative assessment of near-infrared fluorescent proteins Nature methods 2023-09-04 [PMID: 37666982]
• Sherry L, Swanson JJ, Grehan K et al. Protease-Independent Production of Poliovirus Virus-like Particles in Pichia pastoris: Implications for Efficient Vaccine Development and Insights into Capsid Assembly Microbiology spectrum 2023-02-14 [PMID: 36507670] (WB)
• Trolle J, McBee RM, Kaufman A et al. Resurrecting essential amino acid biosynthesis in mammalian cells eLife 2022-09-27 [PMID: 36165439] (WB, Chinese Hamster)
• Niemann P, Schiffer M, Malan D Et al. Generation and Characterization of an Inducible Cx43 Overexpression System in Mouse Embryonic Stem Cells Cells 2022-02-25 [PMID: 35203340] (ICC/IF)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Tominaga K, Tominaga N, Williams EO Et al. 4-Phenylbutyrate restores localization and membrane repair to human dysferlin mutations iScience 2022-01-14 [PMID: 35028538]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Figley MD, Gu W, Nanson JD et al. SARM1 is a metabolic sensor activated by an increased NMN/NAD(+) ratio to trigger axon degeneration Neuron 2021-03-03 [PMID: 33657413]
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Rawson JMO, Duchon A, Nikolaitchik OA et al. Development of a Cell-Based Luciferase Complementation Assay for Identification of SARS-CoV-2 3CL(pro) Inhibitors Viruses 2021-01-27 [PMID: 33498923] (WB, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Huang D, Tran JT, Olson A et al. Vaccine elicitation of HIV broadly neutralizing antibodies from engineered B cells Nat Commun 2020-11-18 [PMID: 33203876] (ELISA, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Szulc DA, Lee XA, Cheng HM, Cheng HM. Bright Ferritin-a Reporter Gene Platform for On-Demand, Longitudinal Cell Tracking on MRI iScience 2020-07-25 [PMID: 32707432] (WB, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Song D, Wang D, Yang Q et al. The lateralization of left hippocampal CA3 during the retrieval of spatial working memory Nat Commun 2020-06-11 [PMID: 32518226] (ICC/IF, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Fonseca JP, Bonny AR, Kumar GR et al. A Toolkit for Rapid Modular Construction of Biological Circuits in Mammalian Cells ACS Synth Biol 2019-11-06 [PMID: 31686495] (WB, ICC/IF, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Bonny A Investigating mechanisms of endogenous, systems-level coordination in stress adaptation and homeostasis Thesis 2020-01-01 (WB, Human)
• Yu FH, Huang KJ, Wang CT. HIV-1 Mutant Assembly, Processing and Infectivity Expresses Pol Independent of Gag Viruses 2020-01-08 [PMID: 31906562] (WB, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Krabbe, S, Paradiso, E Et al. Adaptive disinhibitory gating by VIP interneurons permits associative learning. Nat Neurosci 2019-11-01 [PMID: 31636447] (WB)
• Markus BM, Bell GW, Lorenzi HA, Lourido S. Optimizing Systems for Cas9 Expression in Toxoplasma gondii mSphere 2019-06-28 [PMID: 31243081] (WB)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Szulc DA, Cheng HM. One-Step Labeling of Collagen Hydrogels with Polydopamine and Manganese Porphyrin for Non-Invasive Scaffold Tracking on Magnetic Resonance Imaging Macromol Biosci 2019-01-16 [PMID: 30645045]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.

Reviews for 2A Peptide Antibody (NBP2-59627) (2)

Average Rating: 4.5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Mouse.

reviewed by: Verified Customer WB Mouse 07/13/2023

Summary

• Application: Western Blot
• Sample Tested: NIH/3T3 whole cell lysate
• Species: Mouse
• Lot: D-6

Comments

• Comments: 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 and the protein expression correlates nicely with the viral titer.

reviewed by: Min-Joon Han WB Human 08/14/2018

Summary

• Application: Western Blot
• Sample Tested: Human fibroblast
• Species: Human
• Lot: A-2

FAQs for 2A Peptide Antibody (NBP2-59627). (Showing 1 - 2 of 2 FAQs).

1. 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.
2. 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.