mCherry Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-25158

Novus Biologicals, part of Bio-Techne
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NBP2-25158
NBP2-25158-0.025ml
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Non-species specific

Cited:

Human, Mouse, Avian - Chicken, Donkey, Insect, Primate - Macaca mulatta (Rhesus Macaque), Rabbit

Applications

Validated:

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry Whole-Mount, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Cited:

Block/Neutralize, ELISA, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry Whole-Mount, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Chicken IgY

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all mCherry Antibodies »

Product Specifications

Immunogen

This mCherry Antibody was developed against full length recombinant protein expressed in and purified from E. coli.

Clonality

Polyclonal

Host

Chicken

Isotype

IgY

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

Immunocytochemistry/ Immunofluorescence: mCherry Antibody [NBP2-25158]

Immunocytochemistry/ Immunofluorescence: mCherry Antibody [NBP2-25158]

Immunocytochemistry/Immunofluorescence: mCherry Antibody [NBP2-25158] - HEK293 cells transfected with mCherry, stained with mCherry antibody and viewed in a confocal microscope. Most HEK293 cells are not transfected so only the nucleus of these cells can be visualized with a blue DNA stain. Cells which are transfected with mCherry are bright red, and staining with the mCherry antibody is shown in Green. The green antibody staining is only seen cells which express mCherry, and the superimposition of the green and red results in an orange signal. Interestingly stronger mCherry staining is seen in the nucleus, possibly due to degradation of some mCherry molecules releasing the low molecular weight mCherry fluorochrome.
Western Blot: mCherry Antibody [NBP2-25158]

Western Blot: mCherry Antibody [NBP2-25158]

Western Blot: mCherry Antibody [NBP2-25158] - Analysis of NBP2-25158. Lane 1. Lysate of HEK293 cells transfected with pFin-EF1-mCherry vector. There is a strong clean band at about 29kDa corresponding to mCherry (predicted molecular weight ~27kDa). Lane 2, lysate of untransfected HEK293 cells show no protein bands. The NBP2-25158 antibody was used at 1:2,000 dilution.
Western Blot: mCherry Antibody [NBP2-25158]

Western Blot: mCherry Antibody [NBP2-25158]

Western Blot: mCherry Antibody [NBP2-25158] - Analysis of HEK293 cell lysates, and recombinant protein solutions using chicken mCherry pAb, dilution 1:2000 (Green). [1] protein standard, [2] HEK293, [3] HEK293 cells transfected with an mCherry-HA constract, [4] pure recombinant mCherry protein (predicted molecular weight ~ 27kDa), [5] pure recombinant GFP protein and [6] HEK293 transfected with a GFP construct. The major band at about 30kDa corresponds to mCherry-HA protein and the slightly larger recombinant form runs at about 33kDa due to presence of a His tag and other vector derived sequence. The mCherry antibody reacts strongly with both mCherry constructs and does not react with GFP protein. The blot was simultaneously probed with mouse HSP60 mAb, dilution 1:10000 (Red), which reveals a band at 60kDa only in the cell lysates.
View 2 more images

Applications for mCherry Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:1000

Immunohistochemistry

1:1000

Western Blot

1:2000-1:5000
Application Notes
This mCherry antibody is useful for ICC/IF and Western Blot, where a band can be seen at ~28 kDa. Use in IHC-P, IHC-WhMt, and ELISA reported in scientific literature (PMID: 24924516, 29889212, and 29791133 respectively). Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 31591156). Use in Immunohistochemistry free floating reported in scientific literature (PMID: 31320449).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using NBP2-25158 in the following applications:

Formulation, Preparation, and Storage

Purification

IgY purified

Formulation

Supplied as concentrated total IgY preparation from egg yolk. Exact concentration of target specific IgY is not quantifiable as the preparation contains both immune IgY specific for the target and also irrelevant, non-immune IgY.

Preservative

0.035% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

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

Additional mCherry Products

Product Documents for mCherry Antibody

Product Datasheets

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

Chicken products cannot be exported to Canada.

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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