![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.](http://images.novusbio.com/fullsize/mCherry-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-25158-img0005.jpg "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.")
| Reactivity | All-NASpecies Glossary |
| Applications | WB, ELISA, ICC/IF, IHC, IHC |
| Clonality | Polyclonal |
| Host | Chicken |
| Conjugate | Unconjugated |
| Immunogen | This mCherry Antibody was developed against full length recombinant protein expressed in and purified from E. coli. |
| Isotype | IgY |
| Clonality | Polyclonal |
| Host | Chicken |
| Purity | IgY purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| 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). |
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| 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. |
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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | 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 |
| Purity | IgY purified |
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reviewed by:
Masahiko Takemura |
Immunohistochemistry (whole mount) | Drosophila | 12/08/2016 |
Summary
Comments
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Secondary Antibodies |
Isotype Controls |
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| Masahiko Takemura 12/08/2016 |
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| Application: | Immunohistochemistry (whole mount) | |
| Species: | Drosophila |
![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.](http://images.novusbio.com/fullsize/mCherry-Antibody-NBP2-25158-img0004.jpg "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] - 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.](http://images.novusbio.com/fullsize/mCherry-Antibody-Western-Blot-NBP2-25158-img0006.jpg "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.")
![Immunocytochemistry/Immunofluorescence: mCherry Antibody [NBP2-25158] - Dox-inducible DN Lats2-IRES-mCherry transgenic embryos were imaged before Dox treatment (top panel) and the same embryo was imaged following 24 hr of Dox live (middle panel) and fixed/stained for lineage markers (bottom panel). A representative embryo is shown for each stage. Live mCherry is shown as an extended focus image, immunofluorescence stainings shown as single plane images. mCherry positive ICMs in mosaic transgenic embryos are circled with a dotted line. Arrow points to a rare ICM cell in a 64 cell stage-induced embryo with weak Cdx2 expression, which also co-expressed an ICM marker. Scale bar: 25 um. n indicates number of transgenic embryos analyzed. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28226240/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/mCherry-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-25158-img0007.jpg "Immunocytochemistry/Immunofluorescence: mCherry Antibody [NBP2-25158] - Dox-inducible DN Lats2-IRES-mCherry transgenic embryos were imaged before Dox treatment (top panel) and the same embryo was imaged following 24 hr of Dox live (middle panel) and fixed/stained for lineage markers (bottom panel). A representative embryo is shown for each stage. Live mCherry is shown as an extended focus image, immunofluorescence stainings shown as single plane images. mCherry positive ICMs in mosaic transgenic embryos are circled with a dotted line. Arrow points to a rare ICM cell in a 64 cell stage-induced embryo with weak Cdx2 expression, which also co-expressed an ICM marker. Scale bar: 25 um. n indicates number of transgenic embryos analyzed. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28226240/) licensed under a CC-BY license.")
![Western blot analysis of HEK293 cell lysates using chicken pAb to mCherry, NBP2-25158, dilution 1:2,000, in green, and rabbit pAb to GAPDH, RPCA-GAPDH, dilution 1:5,000, in red: [1] protein molecular weight standard, [2] untransfected HEK293 control cells, [3] HEK293 cells transfected with pCI-Neo-mod vector expressing two tdTomato protein domains, [4] HEK293 cells transfected with pCI-Neo-mod vector expressing one mCherry-HA protein domain, and [5] HEK293 cells transfected with pCI-Neo-mod vector expressing one GFP domain. NBP2-25158 recognizes tdTomato and mCherry proteins revealing major bands at about 60kDa and 30kDa, but does not recognize GFP. The red band at 37kDa corresponds to GAPDH protein here used as a loading control.](http://images.novusbio.com/fullsize/nbp2-25158_chicken-polyclonal-mcherry-antibody-western-blot-264202414221..jpg "Western blot analysis of HEK293 cell lysates using chicken pAb to mCherry, NBP2-25158, dilution 1:2,000, in green, and rabbit pAb to GAPDH, RPCA-GAPDH, dilution 1:5,000, in red: [1] protein molecular weight standard, [2] untransfected HEK293 control cells, [3] HEK293 cells transfected with pCI-Neo-mod vector expressing two tdTomato protein domains, [4] HEK293 cells transfected with pCI-Neo-mod vector expressing one mCherry-HA protein domain, and [5] HEK293 cells transfected with pCI-Neo-mod vector expressing one GFP domain. NBP2-25158 recognizes tdTomato and mCherry proteins revealing major bands at about 60kDa and 30kDa, but does not recognize GFP. The red band at 37kDa corresponds to GAPDH protein here used as a loading control.")
