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.

Western Blot: mCherry Antibody (1C51) [NBP1-96752] - Threonine 276 is required for the intramolecular interaction, and for inhibition of membrane binding. Mutation of threonine 276 to aspartic acid promotes the association of cytohesin 2 with membranes. MDCK cells were transfected with constructs encoding mCherry-tagged wild-type or T276D cytohesin 2 and fractionated into cytosol and total membranes. The fractions were Western blotted with mouse anti-mCherry, mouse anti-E-cadherin and mouse anti-actin. Image collected and cropped by CiteAb from the following publication (doi.org/10.1371/journal.pone.0082084) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: mCherry Antibody (1C51) [NBP1-96752] - HEK293 cells transfected with mCherry and visualized in red. The cells were stained with NBP1-96752 in the green channel, and visualized using a confocal microscope. Transfected cells are yellow, showing overlap of the mCherry and NBP1-96752. Untransfected HEK293 cells do not express Cherry and do not stain with the antibody, but their nuclei can be visualized using a DNA stain (blue).

Western Blot: mCherry Antibody (1C51) [NBP1-96752] - WB assay of the crude extract of HEK293 cells transfected with pFin-EF1-mCherry vector (lane +) and an equal amount of protein extract from untransfected HEK293 cells (lane -). NBP1-96752 binds a major band running at about 28 kDa (observed molecular weight) corresponding to intact full-length mCherry. The two other bands are clearly processed forms of mCherry as they are not present in non-transfected HEK293 cells.

Immunohistochemistry-Frozen: mCherry Antibody (1C51) [NBP1-96752] - Mouse Bone Marrow Sections (Femur). Fixed-frozen and decalcified. tdTomato reporter transgenic mice. tdTomato in hematopoietic cells were detected by anti-mCherry antibody. Antibody is cross-reactive and works well for fixed-frozen bone marrow. Background is low. IHC-Fr image submitted by a verified customer review.

• Reactivity: All-NA
• Applications: WB, Flow, ICC/IF, IHC, IP, KO, Single-Cell Western
• Clone: 1C51
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

mCherry Antibody (1C51) Summary

• 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.
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:500
  • Immunohistochemistry 1:500
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Knockout Validated
  • 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).
• 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: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% glycerol
• Preservative: 5mM Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for mCherry Antibody (1C51)

• 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.

Publications for mCherry Antibody (NBP1-96752)(75)

Publications using NBP1-96752

• Kim H, Lee Y, Lee S, Park B HCMV-encoded viral protein US12 promotes autophagy by inducing autophagy flux Biochemical and Biophysical Research Communications 2023-03-01 [PMID: 36898229] (IP)
• Seth A, Bournat JC, Medina-Martinez O et al. Loss of WNT4 in the gubernaculum causes unilateral cryptorchidism and fertility defects Development (Cambridge, England) 2022-12-01 [PMID: 36448532] (IHC-P)
  
  Details:
  Dilution used 1:500
• Israel S, Drexler Hca, Fuellen G, Boiani M The COP9 signalosome subunit 3 is necessary for early embryo survival by way of a stable protein deposit in mouse oocytes Molecular human reproduction 2021-07-15 [PMID: 34264319]
• Glykofrydis F, Cachat E, Berzanskyte I et al. Bioengineering Self-Organizing Signaling Centers to Control Embryoid Body Pattern Elaboration ACS synthetic biology 2021-05-21 [PMID: 34019395]
• Finke M, Brecht D, Stifel J et al. Efficient splicing-based RNA regulators for tetracycline-inducible gene expression in human cell culture and C. elegans Nucleic acids research 2021-04-24 [PMID: 33893804]
• Peterson J, Li S, Kaltenbrun E et al. Expression of transgenes enriched in rare codons is enhanced by the MAPK pathway Scientific reports 2020-12-17 [PMID: 33335127] (IP, FLOW, WB)
• Singh SR, Meyer-Jens M, Alizoti E et al. A high-throughput screening identifies ZNF418 as a novel regulator of the ubiquitin-proteasome system and autophagy-lysosomal pathway Autophagy 2020-12-27 [PMID: 33249983] (WB)
• Seervai RNH, Jangid RK, Karki M et al. The Huntingtin-interacting protein SETD2/HYPB is an actin lysine methyltransferase Sci Adv 2020-10-01 [PMID: 33008892] (WB, IP)
• Otte CG, Fortuna TR, Mann JR et al. Optogenetic TDP-43 nucleation induces persistent insoluble species and progressive motor dysfunction in vivo Neurobiol. Dis. 2020-09-11 [PMID: 32927062] (WB)
• Begovich K, Vu AQ, Yeo G, Wilhelm JE Conserved metabolite regulation of stress granule assembly via AdoMet J. Cell Biol. 2020-08-03 [PMID: 32609300] (WB)
• Christgen S, Zheng M, Kesavardhana S et al. Identification of the PANoptosome: A Molecular Platform Triggering Pyroptosis, Apoptosis, and Necroptosis (PANoptosis) Front Cell Infect Microbiol 2020-05-29 [PMID: 32547960] (KO, WB)
• Jacomin AC, Petridi S, Di Monaco M et al. Regulation of Expression of Autophagy Genes by Atg8a-Interacting Partners Sequoia, YL-1, and Sir2 in Drosophila Cell Rep 2020-05-26 [PMID: 32460019]
• Batabyal S, Gajjeraman S, Tchedre K et al. Near-Infrared Laser-Based Spatially Targeted Nano-enhanced Optical Delivery of Therapeutic Genes to Degenerated Retina Mol Ther Methods Clin Dev 2020-06-12 [PMID: 32355865] (WB, Human)
• Ma X, Zhu P, Ding Y et al. Retinoid X receptor alpha is a spatiotemporally predominant therapeutic target for anthracycline-induced cardiotoxicity Sci. Adv. 2020-01-01 [PMID: 32064346] (WB, Zebrafish)
• Zhou Y, Wang X, Song M et al. A secreted microRNA disrupts autophagy in distinct tissues of Caenorhabditis elegans upon ageing Nat Commun 2019-10-23 [PMID: 31645592] (WB)
• Ghosh R, Smith S, Nwangwa E et al. Panax quinquefolius (North American ginseng) cell suspension culture as a source of bioactive polysaccharides: Immunostimulatory activity and characterization of a neutral polysaccharide AGC1 International Journal of Biological Macromolecules 2019-07-01 [PMID: 31376448] (WB)
• Fan K, Gravemeyer J, Ritter C et al. MCPyV Large T antigen induced atonal homolog 1 (ATOH1) is a lineage-dependency oncogene in Merkel cell carcinoma J. Invest. Dermatol. 2019-07-05 [PMID: 31283928]
• Kitamata M, Hanawa-Suetsugu K et al. Membrane-Deformation Ability of ANKHD1 Is Involved in the Early Endosome Enlargement. iScience 2019-07-26 [PMID: 31255983] (ICC/IF)
• Min J, Wright WE, Shay JW Clustered telomeres in phase-separated nuclear condensates engage mitotic DNA synthesis through BLM and RAD52 Genes Dev. 2019-07-01 [PMID: 31171703]
• Choi, E;Kikuchi, S;Gao, H;Brodzik, K;Nassour, I;Yopp, A;Singal, AG;Zhu, H;Yu, H; Mitotic Regulators and the SHP2-MAPK Pathway Promote Insulin Receptor Endocytosis and Feedback Regulation of Insulin Signaling. Nat Commun. [PMID: 30931927] (ICC/IF, Mouse)
• Dlugosz Paula, Tresky Roland, Nimpf Johannes Differential Action of Reelin on Oligomerization of ApoER2 and VLDL Receptor in HEK293 Cells Assessed by Time-Resolved Anisotropy and Fluorescence Lifetime Imaging Microscopy. Frontiers in Immunology 2019-02-26 [PMID: 30873003] (ICC/IF, WB, Rat)
• Xu Y, He Z, Song M et al. A microRNA switch controls dietary restriction-induced longevity through Wnt signaling EMBO Rep. 2019-03-14 [PMID: 30872315] (WB, Human)
• Korin B, Rolls A. Application of Chemogenetics and Optogenetics to Dissect Brain-Immune Interactions Methods Mol. Biol. 2018-04-30 [PMID: 29705849] (IF/IHC)
• Plona KL Exploring molecular pathogenesis to streamline future therapeutics in rare diseases using GSD1a as a model Thesis 1905-07-13
• Rieskamp JD, Rosado-Burgos I, Christofi JE et al. Excitatory amino acid transporter 1 supports adult hippocampal neural stem cell self-renewal iScience 2023-07-21 [PMID: 37534178] (IHC-Fr)
  
  Details:
  Dilutions: 1:500
• Park J, Wu Y, Shao W et al. Poly(GR) interacts with key stress granule factors promoting its assembly into cytoplasmic inclusions Cell Reports 2023-08-29 [PMID: 37471224]
• Cmentowski V, Ciossani G, d'Amico E et al. A mechanism that integrates microtubule motors of opposite polarity at the kinetochore corona bioRxiv 2023-09-01 [PMID: 37163019] (WB)
• Sadhu L, Tsopoulidis N, Hasanuzzaman M et al. ARPC5 isoforms and their regulation by calcium-calmodulin-N-WASP drive distinct Arp2/3-dependent actin remodeling events in CD4 T cells eLife 2023-05-10 [PMID: 37162507] (ICC/IF, WB)
• Han B, Gulsevin A, Connolly S et al. Structural analysis of the P132L disease mutation in caveolin-1 reveals its role in assembly of oligomeric complexes The Journal of biological chemistry 2023-03-02 [PMID: 36870682] (ICC/IF)
• Sterling FR, D'Amico J, Brumfield AM et al. StARD9 is a novel lysosomal kinesin required for membrane tubulation, cholesterol transport and Purkinje cell survival Journal of cell science 2023-03-01 [PMID: 36861884] (ICC/IF, Mouse)
• Ghochani Y, Muthukrishnan SD, Sohrabi A et al. A molecular interactome of the glioblastoma perivascular niche reveals integrin binding sialoprotein as a mediator of tumor cell migration Cell reports 2022-10-18 [PMID: 36261010]
• Tian F, Cheng Y, Zhou S et al. Core transcription programs controlling injury-induced neurodegeneration of retinal ganglion cells Neuron 2022-06-24 [PMID: 35767995] (IF/IHC)
• Zhang Y, Liang P, Yang L et al. Functional coupling between TRPV4 channel and TMEM16F modulates human trophoblast fusion eLife 2022-06-07 [PMID: 35670667]
• Willis SD, Hanley SE, Doyle SJ et al. Cyclin C-Cdk8 Kinase Phosphorylation of Rim15 Prevents the Aberrant Activation of Stress Response Genes Frontiers in Cell and Developmental Biology 2022-03-31 [PMID: 35433688] (B/N, WB)
• Bhattacharjee A, UrmOsi A, Jipa A et al. Loss of ubiquitinated protein autophagy is compensated by persistent cnc/NFE2L2/Nrf2 antioxidant responses Autophagy 2022-02-20 [PMID: 35184662]
• Smeeton J, Natarajan N, Anderson T et al. Regeneration of Jaw Joint Cartilage in Adult Zebrafish Frontiers in Cell and Developmental Biology 2022-01-20 [PMID: 35127702]
• Belyaeva V, Wachner S, Gyoergy A et al. Fos regulates macrophage infiltration against surrounding tissue resistance by a cortical actin-based mechanism in Drosophila PLoS biology 2022-01-01 [PMID: 34990456] (WB)
• Eldridge MJG, Hamon MA. Histone H3 deacetylation promotes host cell viability for efficient infection by Listeria monocytogenes PLOS Pathogens 2021-12-20 [PMID: 34929015] (ICC/IF)
• Martinez D, Zhu M, Guidry JJ et al. Mask, the Drosophila ankyrin repeat and KH domain-containing protein, affects microtubule stability Journal of Cell Science 2021-10-15 [PMID: 34553767] (WB)
• Plona K, Eastman J, Drumm M Classifying molecular phenotypes of G6PC variants for pathogenic properties and to guide therapeutic development JIMD Rep 2021-07-14 [PMID: 34258141]
• Walstein K, Petrovic A, Pan D et al. Assembly principles and stoichiometry of a complete human kinetochore module Science Advances 2021-07-02 [PMID: 34193424] (WB)
• Torres JLN, Shanahan SL, Chassefeyre R, Bueno SL LC3B phosphorylation regulates FYCO1 binding and directional transport of autophagosomes Curr Biol 2021-06-19 [PMID: 34146484]
• Koppens M, Davis H, Valbuena G, et al. BMP pathway antagonism by Grem1 regulates epithelial cell fate in intestinal regeneration Gastroenterology 2021-04-05 [PMID: 33819486]
• Sobu Y, Wawro PS, Dhekne HS, Pfeffer SR Pathogenic LRRK2 regulates ciliation probability upstream of Tau Tubulin kinase 2 Proc Natl Acad Sci U S A 2021-03-03 [PMID: 33653948]
• Kang MH, van Lieshout LP, Xu L et al. A lung tropic AAV vector improves survival in a mouse model of surfactant B deficiency Nat Commun 2020-08-09 [PMID: 32764559]
• Jacomin A, Petridi S, Di Monaco M, et al. Self-Regulation of Autophagy Genes Expression by Atg8a and Its Interacting Partners YL-1, Sequoia and Sir2 In Drosophila SSRN Journal 2019-12-05 [PMID: 32460019]
• Wurmthaler LA, Sack M, Gense K et al. A tetracycline-dependent ribozyme switch allows conditional induction of gene expression in Caenorhabditis elegans Nat Commun. 2019-01-29 [PMID: 30700719] (WB)
• Tusco R, Jacomin AC, Jain A et al. Kenny mediates selective autophagic degradation of the IKK complex to control innate immune responses Nat Commun 2017-11-02 [PMID: 29097655] (WB, Drosophila)
• Rebman JK Cellular and Molecular Mechanisms of Collective Migration in Facial Branchiomotor Neurons Thesis (IF/IHC, Zebrafish)
• Gaynor AS, Chen W. Induced prodrug activation by conditional protein degradation J. Biotechnol. 2017-10-20 [PMID: 28912080] (WB)
• Hu Q, Xu Y, Song M et al. BLMP-1 Controls the Timing of Dietary Restriction-Induced Response in Caenorhabditis elegans Caenorhabditis elegans 2023-07-24
• Weir NR, Kamber RA, Martenson JS, Denic V. The AAA protein Msp1 mediates clearance of excess tail-anchored proteins from the peroxisomal membrane Elife 2017-09-14 [PMID: 28906250] (WB)
• Zhu M, Zhang S, Tian X, Wu C. Mask mitigates MAPT- and FUS-induced degeneration by enhancing autophagy through lysosomal acidification Autophagy 2017-08-14 [PMID: 28806139] (WB, Insect)
• Fan Y, Evans CR, Barber KW et al. Heterogeneity of Stop Codon Readthrough in Single Bacterial Cells and Implications for Population Fitness Mol. Cell 2017-08-01 [PMID: 28781237] (WB)
• Jackson CE, Scruggs BS, Schaffer JE, Hanson PI. Effects of Inhibiting VPS4 Support a General Role for ESCRTs in Extracellular Vesicle Biogenesis. Biophys. J. 2017-09-19 [PMID: 28629620] (IF/IHC, Human)
• Tan SLW, Chadha S, Liu Y et al. A Class of Environmental and Endogenous Toxins Induces BRCA2 Haploinsufficiency and Genome Instability. Cell 2017-06-01 [PMID: 28575672] (WB, Human)
• Pan D, Klare K, Petrovic A et al. CDK-regulated dimerization of M18BP1 on a Mis18 hexamer is necessary for CENP-A loading Elife 2017-01-06 [PMID: 28059702] (WB, Human)
• Rebman JK, Kirchoff KE, Walsh GS. Cadherin-2 Is Required Cell Autonomously for Collective Migration of Facial Branchiomotor Neurons. PLoS ONE 2016-10-07 [PMID: 27716840] (IHC-P, Zebrafish)
• Cho A, Kato M, Whitwam T et al. An Atypical Tropomyosin in Drosophila with Intermediate Filament-like Properties Cell Rep 2016-07-26 [PMID: 27396338] (IF/IHC)
• Li X, Rydzewski N, Hider A et al. A molecular mechanism to regulate lysosome motility for lysosome positioning and tubulation. Nat Cell Biol 2016-04-01 [PMID: 26950892] (WB)
• Estruch SB, Graham SA, Deriziotis P, Fisher SE The language-related transcription factor FOXP2 is post-translationally modified with small ubiquitin-like modifiers. Sci Rep 2016-02-12 [PMID: 26867680] (WB, Human)
• Ruas M, Davis LC, Chen CC et al. Expression of Ca2+-permeable two-pore channels rescues NAADP signalling in TPC-deficient cells EMBO J. 2015-04-14 [PMID: 25872774] (ICC/IF, Mouse)
• Zhu M, Li X, Tian X, Wu C. Mask loss-of-function rescues mitochondrial impairment and muscle degeneration of Drosophila pink1 and parkin mutants Hum. Mol. Genet. 2015-03-05 [PMID: 25743185] (WB)
  
  Details:
  Mouse anti-mCherry antibody (clone 1C51) used for WB analysis of mCherry-ATG8 levels in larval muscles expressing 24B-Gal4-driven UAS-mCherry-ATG8 together with the control RNAi, the mask RNAi or UAS-Mask, at 93 h after egg laying. The antibody was used at 1 : 1000 dilution and the data image shows free mCherry as well as ATG8 bound mCherry (Figure 6B).
• Han B, Tiwari A, Kenworthy AK. Tagging strategies strongly affect the fate of overexpressed caveolin-1. Traffic 2015-04-01 [PMID: 25639341]
• Gainullin V G, Hopp K et al. Polycystin-1 maturation requires polycystin-2 in a dose-dependent manner. J Clin Invest 2015-01-02 [PMID: 25574838] (WB)
• Yousef H, Morgenthaler A, Schlesinger C et al. Age-Associated Increase in BMP Signaling inhibits Hippocampal Neurogenesis. Stem Cells. 2014-12-23 [PMID: 25538007]
• Andrews OE,Cha DJ,Wei C, Patton JG. RNAi-Mediated Gene silencing in Zebrafish Triggered by Convergent Transcription Sci Rep. 2014-06-09 [PMID: 24909225] (WB)
  
  Details:
  mCherry antibody (clone 1C51) used in WB - At 72?hpf, GFP+ embryos were de-yolked and protein lysates were prepared from 30 embryos in lysis buffer. 25?ug of total protein was loaded per lane, Lanes 1 and 2 correspond to UIC embryos from 2 separate clutches, Lane 3 (empty), Lane 4: CT-mCherry injected embryos, Lane 5: CT-mCherry + 2.5ng Dicer MO, Lane 6: non-CT mCherry injected embryos. Alpha-tubulin was detected at 50 kDa whereas the band at ~27kDa corresponds to mCherry (Supplemental Figure 1; cropped image in Figure 2C ).
• Liang Hongqing, Esposito Alessandro, De Siddharth et al. Homeostatic control of polo-like kinase-1 engenders non-genetic heterogeneity in G2 checkpoint fidelity and timing. Nat Commun. 2014-06-04 [PMID: 24893992] (WB)
• Liang S, Varrecchia M, Ishida K, Jolly ER. Evaluation of schistosome promoter expression for transgenesis and genetic analysis. PLoS ONE. 2014-05-26 [PMID: 24858918] (WB)
• Li Y, Low LH, Putz U et al. Rab5 and Ndfip1 are involved in Pten ubiquitination and nuclear trafficking. Traffic. 2014-05-05 [PMID: 24798731] (WB)
• Ahier A, Jarriault S. Simultaneous Expression of Multiple Proteins Under a Single Promoter in Caenorhabditis elegans via a Versatile 2A-Based Toolkit. Genetics 2013-12-20 [PMID: 24361941] (WB)
• Hiester KG, Santy LC. et al. The Cytohesin Coiled-Coil Domain Interacts with Threonine 276 to Control Membrane Association. PLoS One. 2013-11-26 [PMID: 24303080] (WB)
• Liang S, Knight M, Jolly ER et al. Polyethyleneimine Mediated DNA Transfection in Schistosome Parasites and Regulation of the WNT Signaling Pathway by a Dominant-Negative SmMef2. PLoS Negl Trop Dis 2013-07-25 [PMID: 23936566] (WB)
• Yamaguchi S, Reid DA, Rothenberg E et al. Changes in Psp protein binding partners, localization and behaviour upon activation of the Yersinia enterocolitica phage shock protein response Mol Microbiol 2013-01-07 [PMID: 23290031] (WB)
• Pan D, Klare K, Petrovic A et al. CDK-regulated dimerization of M18BP1 on a Mis18 hexamer is necessary for CENP-A loading. Elife. 2017-01-06 [PMID: 2805970] (WB, Human)

Reviews for mCherry Antibody (NBP1-96752) (3)

Average Rating: 4.3

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Mouse.

reviewed by: Verified Customer WB Human 04/19/2019

Summary

• Application: Western Blot
• Sample Tested: 293T lysate
• Species: Human
• Lot: 062018

Comments

• Comments: mCherry under a CMV promoter was tested by standard western blot at a 1:1000 dilution in 5% milk. Blot showed good bands with no non-specific bands. Note: This antibody does not detect TdTomato by Western Blot.

reviewed by: Verified Customer Immunofluorescence - fixed-frozen Mouse 10/21/2018

Summary

• Application: Other
• Sample Tested: bone marrow
• Species: Mouse
• Lot: I don't have it any more

Comments

• Comments: Whole Mouse Bone Marrow Sections (Femur) - Fixed-frozen and decalcified. tdTomato reporter transgenic mice

reviewed by: Verified Customer WB Human 03/17/2017

Summary

• Application: Western Blot
• Sample Tested: HeLa transfected with mCherry containing compound,HeLa transfected with mCherry-tagged compound,Hela whole cell lysate
• Species: Human

FAQs for mCherry Antibody (NBP1-96752). (Showing 1 - 6 of 6 FAQs).

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.
2. Would this antibody detect DsRed?
   • NBP1-45840, NBP1-97373 and NBP1-97371 will all recognize DsRed. I have no information about its cross-reactivity of DsRed with NBP1-96752.
3. Do you have any data on the use of NBP1-96752 for immunohistochemistry?
   • 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.
4. Regarding RFP Antibody (NBP1-97373). Does it recognize dTomato?
   • 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.
5. Regarding the mCherry antibody (NBP1-96752) I would like to know if it also detects RFP
   • 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.
6. 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?
   • 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.