Human mCherry Antibody

Catalog #: MAB11041 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB11041-100
MAB11041-SP

Detection of Human mCherry by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human mCherry Antibody Summary

Species Reactivity

Human

Specificity

Detects human mCherry in direct ELISAs.

Source

Recombinant Monoclonal Rabbit IgG Clone # 2783C

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Human mCherry synthetic peptide

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.25 µg/mL

HEK293 human embryonic kidney cell line transfected with mVISTA

Simple Western

25 µg/mL

HEK293 human embryonic kidney cell line transfected with mVISTA

Immunocytochemistry

3-25 µg/mL

Immersion fixed HEK293 human embryonic kidney cell line transfected with mCherry

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells

HEK293 human embryonic kidney cell line transfected with mCherry

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.

Scientific Data

Western Blot

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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.

Immunocytochemistry

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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.

Intracellular Staining by Flow Cytometry

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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.

Western Blot

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Detection of Human mCherry by Simple Western^TM. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

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.

Background: mCherry

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.

References

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.

Long Name

mCherry

Alternate Names

DSRED; mCherry; red fluorescent protein mCherry; Red Fluoroscent Protein