Product DescriptiongoogleChicken anti-mCherry Polyclonal Antibody (Unconjugated), suitable for WB, ICC.
Application(s)ICC, WB
Antibody HostChicken
Antibody TypePolyclonal
SpecificityThe antibody reacts with a band at ~28-30 kDa corresponding to intact full-length mCherry by Western blot on HEK293 cells transfected with mCherry vector. It has also been used successfully for immunocytochemistry.
Species ReactivitySpecies Independent
Immunogen DescriptionRecombinant full length mCherry.
Product DescriptionChicken anti-mCherry Polyclonal Antibody (Unconjugated), suitable for WB, ICC.
Application(s)ICC, WB
Application DetailsWestern Blotting (WB) and Immunocytochemistry (ICC). A dilution of 1:2,000 - 1:5,000 isC recommended for WB. A dilution of 1:500-1,000 is recommended for IC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetmCherry
SpecificityThe antibody reacts with a band at ~28-30 kDa corresponding to intact full-length mCherry by Western blot on HEK293 cells transfected with mCherry vector. It has also been used successfully for immunocytochemistry.
Target Host SpeciesSpecies Independent
Species ReactivitySpecies Independent
Antibody HostChicken
Antibody TypePolyclonal
Antibody IsotypeIgY
ConjugateUnconjugated
Immunogen DescriptionRecombinant full length mCherry.
Purity DescriptionConcentrated purified IgY
FormatLyophilized, without preservatives.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsStore lyophilized, unopened vial at 2-8°C or lower. After reconstitution, prepare aliquots and store at -20°C to -80°C for a higher stability. Avoid freeze-thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundmCherry is an engineered derivative of one of a family of proteins originally isolated from Cnidarians (jelly fish, sea anemones and corals). The mCherry protein was derived from DsRed, a red fluorescent protein from so-called disc corals of the genus Discosoma. DsRed is a 223 amino acid ~28 kDa protein similar in size and properties to GFP, but, obviously, produces a red rather than a green fluorochrome. The original DsRed was engineered extensively in the Tsien lab to prevent it from forming tetramers and dimers and to modify and improve the spectral properties (1-3). The resulting monomeric protein is useful for applications such as Foerster Resonance Energy Transfer (FRET, also known as Fluorescence Resonance Energy Transfer). Several further cycles of mutation, directed modification and evolutionary selection produced mCherry, which is monomeric and has an excitation maximum at 587 nm and and emission maximum at 610 nm (4).
A: Western blot analysis of HEK293 cells transfected with pFin-EF1-mCherry vector. C-1655-100 (1:2,000 dilution) reveals a strong protein band in transfected cells (Lane 1). Non-transfected control cells (Lane 2) show no mCherry protein band. B: Immunofluorescence analysis of mCherry-transfected HEK293 cells with C-1655-100. Specificity of C-1655-100 for mCherry is shown by superimposing mCherry fluorescence (red) with C-1655-100 antibody staining (green), which results in a yellow color. mCherry staining is also seen in the nucleus, which might be due to degradation of mCherry molecules, causing the release of the low molecular weight mCherry fluorochrome.
Left: HEK293 cells transfected with a mCherry (red) were stained with chicken antibody to mCherry (green). Blue: DAPI nuclear stain. Transfected cells appear bright red, and co-stain for mCherry chicken antibody. Superimposition of both signals results in yellow-orange colour. Right: Western blot analysis of transfected and control HEK293 cell lysates, and recombinant proteins with chicken antibody to mCherry (1:2,000, green). [1] protein standard, [2] HEK293 control, [3] HEK293 cells transfected with mCherry, [4] recombinant mCherry protein, [5] recombinant GFP protein, [6] HEK293 transfected with GFP. The major band at about 30 kDa corresponds to mCherry protein and the slightly larger recombinant form runs at about 33 kDa due to presence of a His tag and other vector derived sequences. No cross-reactivity with GFP protein is observed. The same blot was simultaneously probed with a mouse antibody to HSP60 (red), which reveals a band at 60 kDa only in cell lysates.
General ReferencesBaird GS, Zacharias DA, Tsien RY. Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. Proc Natl Acad Sci U S A. 97:11984-9 (2000). Gross LA, Baird GS, Hoffman RC, Baldridge KK, Tsien RY. The structure of the chromophore within DsRed, a red fluorescent protein from coral. Proc Natl Acad Sci U S A. 97:11990-5 (2000). Heikal AA, Hess ST, Baird GS, Tsien RY, Webb WW. Molecular spectroscopy and dynamics of intrinsically fluorescent proteins: coral red (dsRed) and yellow (Citrine). Proc Natl Acad Sci U S A. 97:11996-2001 (2000). Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature Biotechnology 22:1567-1572 (2004).
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