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Key Product Details

Species Reactivity

Non-species specific

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockout Validated, Single Cell Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 1C51

Concentration

1 mg/ml

Product Summary for mCherry Antibody (1C51)

Immunogen

This mCherry Antibody (1C51) was developed against recombinant full-length mCherry purified from E. coli.

Specificity

This mCherry Antibody (1C51) does not cross react with GFP.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

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.

Scientific Data Images for mCherry Antibody (1C51)

Immunohistochemistry: mCherry Antibody (1C51) [NBP1-96752] - Cadherin-2 is required cell autonomously for caudal migration of FBMNs. (A-I) Whole-mount immunocytochemistry showing dorsal views of Tg (isl1:GFP) (A-C) and Tg (isl1:cdh2_EC-mCherry)vc25 transgenic embryos (D-I) at 38 hpf embryos. Embryos are labeled with a-GFP (green) (A,D,G) and a-mCherry (red) (B,E,H) antibodies. (A-C) Wild-type Tg (isl1:GFP) embryos with FBMNs fully migrated into r6. (D-I) Defective caudal migration of FBMNs in Tg (isl1:GFP)/Tg(isl1:cdh2_EC-mCherry) vc25 embryos carrying one copy of the transgene (hemizygous) or two copies (homozygous). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0164433) licensed under a CC-BY license.
Western Blot: mCherry Antibody (1C51) [NBP1-96752] - Analysis of HEK293 cell lysates and recombinant protein solutions using mCherry antibody, dilution 1:1,000 (Green). [1] protein standard, [2] HEK293, [3] HEK293 cells transfected with mCherry-HA construct, [4] mCherry recombinant protein, [5] GFP recombinant protein, and [6] HEK293 transfected with GFP construct. Major band at about 30 kDa corresponds to mCherry protein (predicted molecular weight: 27 kDa). mCherry antibody does not react with GFP protein. The same blot was simultaneously probed with chicken HSP60 pAb, dilution 1:5,000 in red which reveals band at 60 kDa seen only in cell lysates.
Western Blot: mCherry Antibody (1C51) [NBP1-96752] - Fluorescent signals and immunoblots of the dual fluorescence reporter of cup-5 32-UTR in WT worms and mir-83(-) mutants at day 1 of adulthood. Quantification is from the western blots. GFP blots and WT worms serve as controls for loading and normalization respectively. Scale bar: 100 i1/4m. n = 3 independent experiments. Image collected and cropped by Citeab from the following publication (A secreted microRNA disrupts autophagy in distinct tissues of Caenorhabditis elegans upon ageing. Nat Commun (2019) licensed under a CC-BY license.

Applications for mCherry Antibody (1C51)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry

1:500

Single Cell Western

100 ug/mL

Western Blot

1:1000 - 1:2000
Application Notes
Use in Flow reported in scientific literature (PMID:33335127). Use in IHC and IHC-P reported in scientific literature (PMID: 27396338 and 27716840 respectively).
mCherry antibody validated for IHC-Frozen from a verified customer review.
Use in Immunoprecipitation reported in scientific literature (PMID: 33008892).
Use in Knockout Validation was reported in scientific literature (PMID: 32547960).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 4.3 using NBP1-96752 in the following applications:

Published Applications

Read 79 publications using NBP1-96752 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

50% PBS, 50% glycerol

Preservative

5mM Sodium Azide

Concentration

1 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: mCherry

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

Long Name

mCherry

Alternate Names

DSRED, red fluorescent protein mCherry, Red Fluoroscent Protein

Product Documents for mCherry Antibody (1C51)

Certificate of Analysis

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

Product Specific Notices for mCherry Antibody (1C51)

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 mCherry Antibody (1C51)

Showing  1 - 5 of 5 FAQs Showing All
  • Q: Would this antibody detect DsRed?

    A: NBP1-45840, NBP1-97373 and NBP1-97371 will all recognize DsRed. I have no information about its cross-reactivity of DsRed with NBP1-96752.

  • Q: Do you have any data on the use of NBP1-96752 for immunohistochemistry?

    A: At this time we do not have any date on the use of NBP1-96752 in Immunohistochemistry. If you would be interested in testing this antibody, I would invite you to take a look at our Innovator's Reward Program.

  • Q: Regarding RFP Antibody (NBP1-97373). Does it recognize dTomato?

    A: I am sorry, but product NBP1-97373 has been discontinued. However, I have now received confirmation from the lab that mCherry Antibody (1C51) NBP1-96752 does indeed bind to dTomato, and so we would recommend this antibody for you.

  • Q: Regarding the mCherry antibody (NBP1-96752) I would like to know if it also detects RFP

    A: We have not tested our mCherry antibody with catalogue number NBP1-96752 against RFP, however since the two proteins share a high degree of sequence homology the antibody is likely to recognise RFP. As we have not performed this testing in house however, we cannot guarantee that NBP1-96752 will or will not cross-react with RFP.

  • Q: I'm looking for an mCherry antibody to use for WB and got a paper that references your NBP1-96752 and was thinking to buy it. But then I saw your WB picture (image 6) in your webpage, and I don't really get why the antibody does not see only one band (about 28kDA) for the mCherry. What are these "different processed forms" supposed to be?

    A: mCherry differs from other GFP-derived proteins by maturing extremely rapidly, and is highly photostable and resists photobleaching. (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2910338/). In short this protein does not naturally occur in the higher mammals, and this is relevant to answer your concern. Since not present in the higher mammals, the only way for our lab to easily assess whether NBP1-96752 was specific to mCharry protein was using transiently transfection of its gene (in pFin-EF1-mCherry vector) into the host of interests, in this case HEK293 cells. Therefore, the crude extract of HEk293 cells transfected with the gene (lane + on the posted WB image) could reveal the mCherry species by the antibody, but an equal amount of protein extract from untransfected HEK293 cells (lane -) could not. In these experiments, it was not the endogenous protein was detected. The transiently expressed protein may be associated with many detection artifacts, namely, degradation easier, pre-maturation termination of transcription/translation, aggression of protein due to over expression of the proteins in situ, and many others. All of these might be contributing to the detection of the mCherry bands, as those see on the image.
    Since this antibody have been cited in many publications (9), 8 of which were using the antibody in WB, we are confident that the quality of NBP1-96752 is good. In fact we 100% guarantee it to produce the positive results in WB. Or we will refund or free replacement of any primary antibody with the similar price.

Showing  1 - 5 of 5 FAQs Showing All
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