EnCor Biotechnology

Recombinant Full Length mCherry Protein Cat# Prot-r-mCherry

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Description

      The mCherry protein is derived from DsRed, a red fluorescent protein from the disc coral genus Discosoma. The protein is 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 lob of Roger Tsien to prevent it from forming tetramers and dimers and to modify and improve the spectral properties. Several further cycles of mutation, directed modification and evolutionary selection produced mCherry. The mCherry protein sequence was deposited in NCBI entry AY678264, identical to that in Uniprot entry X5DSL3, and is also identical to that found in many expression vectors such as pGGD003.

Amount: 50µg
Amount: 50µg
      The lane on the left contains Biorad SDS-PAGE molecular weight standards of the indicated molecular size. His-tagged recombinant mCherry was run out on an SDS-PAGE gel at 3µg in the second lane. BSA was also run at 9µg, 6µg and 3µg in the next lanes as indicated. The vector adds an C-terminal His-tag which was use to purify the protein and this adds about 5kDa to the molecule, which therefore runs at about 34kDa. The mCherry protein is known to be somewhat unstable, and this accounts for the band at about 20kDa, which is clearly derived from the intact protein as it is immunoreactive with mCherry antibodies.

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Name: Recombinant mCherry protein, Cat# Prot--r-mCherry
HGNC Name: NA
UniProt: X5DSL3 (X5DSL3_ANAMA)
RRID: NA
Format: 1mg/mL in 6M urea, 10mM Phosphate pH=7.5
Applications: Fluorescent protein standard, immunogen for antibody production
Storage: Stable at 4°C for several months. For longer term store at -20°C or lower

      The mCherry protein is derived from DsRed, a red fluorescent protein from the disc coral genus Discosoma. The protein is 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 Roger Tsien lab to prevent it from forming tetramers and dimers and to modify and improve the spectral properties. Several further cycles of mutation, directed modification and evolutionary selection produced mCherry. The mCherry protein sequence was deposited in NCBI entry AY678264, identical to that in Uniprot entry X5DSL3, and is also identical to that found in many expression vectors such as pGGD003. We generated a cDNA encoding full length mCherry protein expressed this in E. coli. The vector adds an C-terminal His-tag which was use to purify the protein and this, along with some other vector derived sequence, adds about 5kDa to the molecule. The construct therefore has a total size of about 34 kDa as shown.

This antibody was made against a recombinant construct expressed in and purified from E. coli using the pET29a (+) vector. This vector adds a few N and C-terminal amino acids which are underlined below. The N terminus contains an S-tag, highlighted in red below, which can be used to purify the protein. However we used the C-terminal His-tag (green below) to purify the protein. The sequence is identical to that found in a series of widely used expression vectors


MKETAAAKFE RQHMDSPDLG TLVPRGSMAD IGSEFMVSKG EEDNMAIIKE FMRFKVHMEG  60
SVNGHEFEIE GEGEGRPYEG TQTAKLKVTK GGPLPFAWDI LSPQFMYGSK AYVKHPADIP 120
DYLKLSFPEG FKWERVMNFE DGGVVTVTQD SSLQDGEFIY KVKLRGTNFP SDGPVMQKKT 180
MGWEASSERM YPEDGALKGE IKQRLKLKDG GHYDAEVKTT YKAKKPVQLP GAYNVNIKLD 240
ITSHNEDYTI VEQYERAEGR HSTGGMDELY KVDKLAAALE HHHHHH 286

Number of amino acids: 286
Molecular weight: 32262.40
Theoretical pI: 5.83

Amino acid composition:
Ala (A) 18 6.3%
Arg (R) 10 3.5%
Asn (N) 7 2.4%
Asp (D) 18 6.3%
Cys (C) 0 0.0%
Gln (Q) 9 3.1%
Glu (E) 28 9.8%
Gly (G) 28 9.8%
His (H) 13 4.5%
Ile (I) 11 3.8%
Leu (L) 17 5.9%
Lys (K) 27 9.4%
Met (M) 13 4.5%
Phe (F) 12 4.2%
Pro (P) 14 4.9%
Ser (S) 15 5.2%
Thr (T) 14 4.9%
Trp (W) 3 1.0%
Tyr (Y) 12 4.2%
Val (V) 17 5.9%


Total number of negatively charged residues (Asp + Glu): 46
Total number of positively charged residues (Arg + Lys): 37

Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.

Ext. coefficient 34380
Abs 0.1% (=1 g/l) 1.066

1. Baird GS, Zacharias DA, Tsien RY. Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. PNAS 97:11984-9 (2000).
2. Gross LA et al. The structure of the chromophore within DsRed, a red fluorescent protein from coral. PNAS 97:11990-5 (2000).
3. Heikal AA et al. Molecular spectroscopy and dynamics of intrinsically fluorescent proteins: coral red (dsRed) and yellow (Citrine). PNAS 97:11996-2001 (2000).
4. Shaner NC et al. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat. Biotech. 22:1567-1572 (2004).

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