mCherry Antibody

Western Blot: mCherry Antibody [NBP2-43727] - Non-transfected (-) and transfected (+) 293T whole cell extracts (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with mCherry antibody diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

Immunoprecipitation: mCherry Antibody [NBP2-43727] - Analysis of 293T whole cell lysate/extractA : 20 ug whole cell lysate/extract of mcherry protein expressing 293T cellsB : Control with 2.5 ug of pre-immune rabbit IgGC : Immunoprecipitation of mcherry protein by 2.5 ug of mcherry antibody 7.5% SDS-PAGE The immunoprecipitated mcherry protein was detected by mcherry antibody diluted at 1 : 3000. EasyBlot anti-rabbit IgG (HRP) was used as a secondary reagent.

• Reactivity: All-NA
• Applications: WB, ICC/IF, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

mCherry Antibody Summary

• Immunogen: Recombinant protein conjugated synthetic peptide encompassing a sequence within the center region of mCherry. The exact sequence is proprietary.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/Immunofluorescence
  • Immunoprecipitation 1:100-1:500
  • Western Blot 1:500-1:3000
• Theoretical MW: 27 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS, 20% Glycerol (pH7).
• Preservative: 0.025% Proclin 300
• Purity: Immunogen affinity purified

Alternate Names for mCherry Antibody

• DSRED
• mCherry
• red fluorescent protein mCherry
• Red Fluoroscent Protein

Background

mCherry is a monomeric red fluorescent protein (mRFP) belonging to the mFruits family which is brighter and more photostable compared to the first-generation mRFP1, making them ideal for fluorescence microscopy (1). mCherry has an excitation maximum at 587 nm and an emission maximum at 610 nm. mCherry protein was derived from DsRed, a red fluorescent protein from the coral Discosoma (disc anemone) (2). The red chromophore of DsRed has a similar topology to GFP, the green fluorescent protein isolated from the jellyfish Aequorea Victoria, but has extended pi-electron conjugation resulting in red-shifted absorbance and emission (3). mCherry is 236 amino acids (aa) in length with a theoretical molecular weight of 28 kDa and has a crystal structure with the chromophore forming a central helix shielded within an eleven-stranded beta-barrel (3).

mCherry can be used as a long-wavelength hetero-FRET (fluorescence resonance energy transfer) acceptor and probe for homoFRET experiments given its high peak molar absorptivity, folding efficiency, and superior spectral properties (4). Additionally, because mCherry does not interfere with other plasmids or alter the growth of Legionella species during intracellular growth, it can be used for constitutive gene expression in a variety of gram-negative bacterial species (5). For example, a plasmid developed to constitutively express mCherry under the Ptac promoter has been used in several Legionella species including L. pneumophila, the causative agent of Legionnaires' disease (5).

References

1. Shaner, N. C., Steinbach, P. A., & Tsien, R. Y. (2005). A guide to choosing fluorescent proteins. Nature Methods, 2(12), 905-909. doi:10.1038/nmeth819

2. Bevis, B. J., & Glick, B. S. (2002). Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed). Nature Biotechnology, 20(1), 83-87. https://doi.org/10.1038/nbt0102-83

3. Wall, M. A., Socolich, M., & Ranganathan, R. (2000). The structural basis for red fluorescence in the tetrameric GFP homolog DsRed. Nature Structural Biology, 7(12), 1133-1138. https://doi.org/10.1038/81992

4. Akrap, N., Seidel, T., & Barisas, B. G. (2010). Forster distances for fluorescence resonant energy transfer between mCherry and other visible fluorescent proteins. Analytical Biochemistry, 402(1), 105-106. https://doi.org/10.1016/j.ab.2010.03.026

5. Gebhardt, M. J., Jacobson, R. K., & Shuman, H. A. (2017). Seeing red; the development of pON.mCherry, a broad-host range constitutive expression plasmid for Gram-negative bacteria. Plos One, 12(3), e0173116. https://doi.org/10.1371/journal.pone.0173116

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.

⚠ WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause reproductive toxicity with developmental effects.  For more information go to www.P65Warnings.ca.gov.

Publications for mCherry Antibody (NBP2-43727)(1)

Publication using NBP2-43727

• Lee A, Rayner SL, Gwee SSL et al. Pathogenic mutation in the ALS/FTD gene, CCNF, causes elevated Lys48-linked ubiquitylation and defective autophagy Cell. Mol. Life Sci. 2017 Aug 29 [PMID: 28852778] (Human)

FAQs for mCherry Antibody (NBP2-43727). (Showing 1 - 1 of 1 FAQ).

1. Does this antibody cross-react with GFP epitopes? As I would like to use both GFP and mCherry antibodies during histochemistry I would not like them to cross-react.
   • mCherry and GFP share just 29% sequence similarity, so this antibody is not predicted to cross-react to GFP and has never shown any ability to detect GFP in testing.