Human mCherry Antibody Summary
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Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human mCherry by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line either mock transfected or transfected with mVISTA. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human mCherry Monoclonal Antibody (Catalog # MAB11041) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for mCherry at approximately 30-80 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
mCherry in HEK293 Human Cell Line Transfected with mCherry. mCherry was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with mCherry using Rabbit Anti-Human mCherry Monoclonal Antibody (Catalog # MAB11041) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of mCherry in HEK293 Human Cell Line transfected with mCherry by Flow Cytometry. HEK293 human cell line transfected with mCherry (filled histogram) or EGFP (open histogram) was stained with Rabbit Anti-mCherry Monoclonal Antibody (Catalog # MAB11041), followed by Allophycocyanin-conjugated Anti-Rabbit IgG F(ab')2Secondary Antibody (F0111). Gates were set based on isotype control antibody (MAB1050, data not shown). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our Staining Intracellular Molecules protocol.
Detection of Human mCherry by Simple WesternTM. Simple Western lane view shows lysates of HEK293 human embryonic kidney cell line either mock transfected or transfected with mVISTA, loaded at 0.2 mg/mL. Specific bands were detected for mCherry at approximately 36 and 110 kDa (as indicated) using 25 µg/mL of Rabbit Anti-Human mCherry Monoclonal Antibody (Catalog # MAB11041). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
mCherry is a member of the mFruits family of monomeric red fluorescent proteins. It was derived from DsRed of Discoscoma sea anemones. mFruit are second-generation monomeric red fluorescent proteins that have improved brightness and photostability compared to first-generation proteins. The gene for mCherry is 711bp long and the protein is made up of 236 residues with a mass of 76.722 kDa. mCherry is used in fluorescence microscopy as an intracellular probe where constitutive gene expression is desired and other experimental approaches require coordinated control of multiple genes.
- Shaner, N.C. Campbell, R.E. Steinbach, P.A. Giepmans B.N.G. Palmer A.E. Tsien, R Y. "Improved Monomeric Red, Orange and Yellow Fluorescent Proteins Derived from Discomsoma Sp. Red Fluorescent Protein" Nature Biotechnology. 22(12):1567. 2004 Nov. 21.
- Shu, X. Shaner N C. Yarbrough, C.A. Tsien, R Y. Remington S.J. "Novel Chromophores and Buried Charges Control Color in mFruits" Biochemistry. 45(32):9639. 2006 Aug.
- Gebhardt, M.J. Jacobson, R.K. Shuman, H. "Seeing Red; The Development of pON mCherry, a Broad-host Range Constitutive Expression Plasmid for Gram-negative Bacteria" POS ONE. 12(3):e173116. 2017 Mar. 3.
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