mcherry Monoclonal Antibody (16D7) Catalog Number M11217 Product data sheet

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1 Website: thermofisher.com Customer Service (US): ext. 1 Technical Support (US): ext. 441 mcherry Monoclonal Antibody (16D7) Catalog Number M11217 Product data sheet Details Size 100 µl Host/Isotope Rat / IgG2a Class Monoclonal Type Antibody Clone 16D7 Immunogen full-length protein mcherry Conjugate Unconjugated Form Liquid Concentration 2 mg/ml Purification purified Storage buffer PBS Contains 0.09% sodium azide Storage Conditions 4 C Species Reactivity Tested species reactivity Published species reactivity Tag Not Applicable Tested Applications Dilution * Flow Cytometry (Flow) Immunocytochemistry (ICC) Immunohistochemistry (IHC) Immunoprecipitation (IP) Western Blot (WB) Published Applications Immunocytochemistry (ICC) Immunohistochemistry - Free Floating (IHC (Free)) Immunoprecipitation (IP) Immunohistochemistry (IHC) * Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.

2 Product Images For mcherry Monoclonal Antibody (16D7) (M11217) in IF Intestinal tissue from a transgenic mouse expressing mcherry in all tissues was isolated and fixed in 4% paraformaldehyde. mcherry Rat Monoclonal Antibody (M11217) was used at a 1:15,000 dilution. Using the ImmPRESS Anti-Rat Ig (peroxidase) Polymer Detection Kit (Vector Laboratories) and following the manufacturer and quote;s instructions, the sections were incubated in peroxidase substrate solution until the desired stain intensity developed. A) Fluorescent image detecting mcherry expression and B) HRPstained image. (M11217) in IF Intestinal tissue from a transgenic mouse expressing mcherry in all tissues was isolated and fixed in 4% paraformaldehyde. mcherry Rat Monoclonal Antibody (M11217) was used at a 1:15,000 dilution. Using the ImmPRESS Anti-Rat Ig (peroxidase) Polymer Detection Kit (Vector Laboratories) and following the manufacturer and quote;s instructions, the sections were incubated in peroxidase substrate solution until the desired stain intensity developed. A) Bright field image, B) HRP-stained image.

3 (M11217) in IF U2OS cells were transduced using an adenoviral construct expressing mcherry. A) Native expression of mcherry detected posttransduction using Texas Red filters (562 nm/624 nm) B) Anti-Cherry antibody added and cells imaged using the Cy5 filter set (628 nm/692 nm) C) mcherry expression detected by adding antimcherry and Alexa Fluor 647 goat anti-rat (Product # A-21247) (M11217) in WB After transfer of the proteins, the nitrocellulose membranes were probed with either (A) rabbit anti- GFP (Product # A-11122) and secondary antibody Alexa Fluor 647 Goat Anti-Rabbit (Product # A ), (B) rat antimcherry (M11217) and secondary antibody Alexa Fluor 647 Goat Anti-Rat (Product # A-21247) or (C) rabbit anti-rfp (Product # R10367) and secondary antibody Alexa Fluor 647 Goat Anti- Rabbit (Product # A ). Lanes 1 and 10: Novex Sharp Pre- Stained Protein Standards (LC5800). Lane 2: 20 µg of U2OS cell lysate expressing plasma membrane targeted TagRFP. Lane 3: mkate RFP-P62 fusion protein. Lane 4: TagGFP- P62 fusion protein. Lane 5: plasma membrane targeted Emerald GFP. Lanes 6 and 7: untargeted mcherry. Lanes 8 and 9: control. The anti-mcherry antibody is specific for mcherry protein and does not cross-react with either GFP or RFP.

4 (M11217) in Flow U2OS cells were transduced with plasma membrane targeted-gfp (light green line), emerald GFP (dark green line), plasma membrane targeted-tagrfp (purple line), P62-mKate (red line), or mcherry (pink line). After trypsinization, cells were fixed and permeablized. They were then blocked with normal rat IgG and labeled with 1 µg of a direct conjugate of anti-mcherry- PacificBlue. Untransduced cells stained with antibody served as a negative control. All proteins were expressed, as determined by microscopy (data not shown). (M11217) in Flow U20S cells expressing mcherry were analyzed using 405 nm excitation and 450/40 nm band pass emission on an Attune Acoustic Focusing Cytometer. The histogram shows cells stained with mcherry rat monoclonal antibody conjugated with Pacific Blue (black line) and unstained cells (gray line).

5 (M11217) in IP Hela cells were transduced with adenovirus-mcherry using the Novex Immunoprecipitation Kit Dynabeads Protein G (Product # 10007D). Samples were analyzed by SDS-PAGE using a NuPAGE 4-12% Bis-Tris Gel. Samples were transferred to nitrocellulose membrane using the iblot Transfer Device. Rat antimcherry antibody was used at 1:1000 dilution with goat anti-rat Alexa Fluor 647 secondary at 1:1000 dilution to detect mcherry in the cell lysate samples. The antimcherry antibody detected mcherry expression (bands at ~30K and ~24K). The 30K band is intact mcherry, while the 24K band represents the mcherry degradation product.lane 1: Novex Sharp Pre- Stained Protein Standards (LC5800). Lane 2: 20 µg U2OS control lysate. Lane 3: 100 ng immunopreciptated purified Hela lysate. Lane 4: 40 µg HeLa lysate supernatant from IPP purification.

6 PubMed References For mcherry Monoclonal Antibody (16D7) 1 Immunocytochemistry References Not Applicable / 1: Immunohistochemistry - Free Floating References Not Applicable / 1: Immunoprecipitation References Not Applicable / Not Cited M11217 was used in immunocytochemistry and western blot to study allorecognition using Botryllus schlosseri. Immunogenetics (Oct 2015; 67: 605) "Molecular evolution and in vitro characterization of Botryllus histocompatibility factor." Author(s):Taketa DA,Nydam ML,Langenbacher AD,Rodriguez D,Sanders E,De Tomaso AW PubMed Article URL: M11217 was used in immunohistochemistry - free floating to develop methods to quantitatively analyze the input-output relationship of neural circuits Nature (Aug 2015; 524: 88) "Viral-genetic tracing of the input-output organization of a central noradrenaline circuit." Author(s):Schwarz LA,Miyamichi K,Gao XJ,Beier KT,Weissbourd B,DeLoach KE,Ren J,Ibanes S,Malenka RC,Kremer EJ,Luo L PubMed Article URL: 1 Immunohistochemistry References M11217 was used in immunoprecipitation to uncover a role for ATG12-ATG3 in late endosome function that is distinct from autophagy. Nature cell biology (Mar 2015; 17: 300) "ATG12-ATG3 interacts with Alix to promote basal autophagic flux and late endosome function." Author(s):Murrow L,Malhotra R,Debnath J PubMed Article URL: M11217 was used in immunohistochemistry to identify the mechanisms underlying the evolution of a uniform pattern in Danio albolineatus Not Applicable / 1:300 Nature communications (Nov 2014; 5: null) "Pigment cell interactions and differential xanthophore recruitment underlying zebrafish stripe reiteration and Danio pattern evolution." Author(s):Patterson LB,Bain EJ,Parichy DM PubMed Article URL:

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