Mouse Cherry Monoclonal Antibody

Cherry (mCherry, DsRed, Discosoma)

Cat. Num. AAA6012446

• Applications: Western Blot, Immunofluorescence
• Purity: Affinity Purified; Purified by Protein G affinity chromatography.
• Synonyms: Cherry; Monoclonal Antibody; Cherry (mCherry; DsRed; Discosoma); Anti -Cherry (mCherry; anti-Cherry antibody

• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG2a
• Clone Number: 13B813
• Specificity: Recognizes mCherry.
• Purity/Purification: Affinity Purified; Purified by Protein G affinity chromatography.
• Form/Format: Supplied as a liquid in 10mM sodium azide.

• Applicable Applications for anti-Cherry antibody: Western Blot (WB), Immunofluorescence (IF)
• Application Notes: Suitable for use in Immunofluorescence and Western Blot.; Dilution: Immunofluorescence: 1:500; Western Blot: 1:2000
• Immunogen: Recombinant full length His tagged mCherry purified from E. coli.
• Preparation and Storage: May be stored at 4 degree C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degree C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Testing Data

(Blot of crude extract of HEK293 cells transfected with pFin-EF1-mCherry vector in lane labelled "+". The "-" lane is a blot of an equal amount of protein extract from untransfected HEK293 cells. MBS6012446 binds a major band running at ~28kD 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.)

Testing Data

(Shows HEK293 cells transfected with mCherry and visualized in red. The cells were stained with MBS6012446 in the green channel, and visualized using a confocal microscope. Transfected cells are yellow, showing overlap of the mCherry and the MBS6012446 antibody. 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). Blot and transfected cells courtesy of the Semple-Rowland lab at the University of Florida.)

Related Product Information for anti-Cherry antibody

mCherry is an engineered derivative of one of a family of proteins originally isolated from Cnidarians (jelly fish, sea anemones and corals). The prototype for these fluorescent proteins is Green Fluorescent Protein (GFP), which is a ~27kD protein isolated originally from the jellyfish Aequoria victoria. GFP was the basis of the 2008 Nobel Prize in Chemistry, awarded to Osamu Shimomura, Martin Chalfie and Roger Tsien, specifically "for the discovery and development of the green fluorescent protein, GFP". On expression from the GFP gene, GFP protein will fold correctly and fluoresce strongly, the development of fluoresence requires no cofactors except molecular oxygen. It can therefore be expressed in fluorescent form in essentially any prokaryotic or eukaryotic cell under aerobic conditions (1). As a result DNA encoding GFP can be fused to DNA encoding other proteins as a means to visualize the resulting fusion protein in live cells or animals. Engineered forms of GFP and relatives have been developed to monitor Calcium levels, protease activation and in a variety of other processes in real time. A whole range of GFP derivatives with different spectral properties have been developed, largely in the Tsien lab. The mCherry protein was derived from DsRed, a red fluorescent protein from so-called disc corals of the genus Discosoma. DsRed is a 223 amino acid ~28kD protein similar in size and properties to GFP, but, obviously, produces a red rather than a green fluorochrome. The original DsRed was engineered extensively in the Tsien lab to prevent it from forming tetramers and dimers and to modify and improve the spectral properties (2-4). The resulting monomeric protein is useful for applications such as Förster Resonance Energy Transfer (FRET, also known as Fluorescence Resonance Energy Transfer). Several further cycles of mutation, directed modification and evolutionary selection produced mCherry, which is monomeric and has an excitation maximum at 587 nm and and emission maximum at 610 nm (5). We expressed the mCherry protein sequence shown in reference 5 in bacteria, purified out the mCherry and raised several mouse monoclonal antibodies. The antibody was found to stain a band of the expected size in Hek293 cells transfected with a vector designed to express mCherry.

Product Categories/Family for anti-Cherry antibody

Antibodies; Abs to Assay Detection

NCBI and Uniprot Product Information

• NCBI GI #: 159162378
• NCBI Official Full Name: Chain A, Solution Structure Of The Major Cherry Allergen Pru Av 1 Mutant E45w