EnCor Biotechnology

Recombinant Human Full Length MAP2C Protein Cat# PROT-r-MAP2C

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Description

      Microtubule associated protein 2 or MAP2 is a major microtubule binding protein of neurons, where it is concentrated in dendrites and perikarya but absent from axons. There is a single mammalian MAP2 gene which may generate multiple lower molecular weight forms usually named MAP2C and MAP2D which run on SDS-PAGE gels at 60-70kDa, though are actually much smaller in molecular size. These forms are found early in development but as the animal matures are replaced by MAP2A and B which are much larger in molecular size. These two forms include the so-called projection domain, a long insert which results in both molecules running at ~240kDa on SDS-PAGE gels. We have expressed five different recombinant forms of MAP2 based on the human sequence which in combination cover almost the entire human MAP2 molecule.
Amount: 50µg
Amount: 50µg
      The S lane shows molecular weight standards of the indicated apparent molecular weights in kiloDaltons. The next 5 lanes show 2µg amounts of purified recombinant forms of various human MAP2 proteins. D is recombinant full length MAP2D and C is recombinant full length MAP2C. MAP2D is 559 amino acids while MAP2C is 471, missing one of the microtubule binding domains of MAP2D and a more N-terminal 57 amino acid N-terminal sequence. Lanes P1, P2 and P3 show each recombinant segments of the human MAP2 projection domain, found in MAP2A and B but absent from MAP2C and D. The P1 construct is amino acids 233-684, the P2 is 712-1136 and P3 is 1137-1588. All of the constructs run more slowly on SDS-PAGE than expected from their molecular size, probably due to their high content of charged amino acids. These constructs were used to make a series of mouse monoclonal antibodies. The lane marked BSA contains 2µg of bovine serum albumin, another gel standard.

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Name: MAP2C, recombinant construct, PROT-r-MAP2C
HGNC Name: MAP2
UniProt: P11137
Molecular Weight: 55.2kDa, ~60kDa by SDS-PAGE
Format: 1mg/mL in 6M Urea
Applications: Protein standard, immunogen, ELISA

      We inserted cDNAs encoding the human MAP2C sequences into the pET29a eukaryotic expression vector. 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 5 kDa to the molecule. This sequence includes an S-tag affinity peptide (red) and an enterokinase cleavage site (green), and the His tag sequence is underlined.

      This is the shortest transcript of the human MAP2 gene, including only 3 of the C-terminal microtubule binding domains. The sequence is identical to that in NP_001350842.1. The construct was expressed in pET29a and has a few N and C-terminal amino acids derived from the vector, including a C-terminal His-tag.


MKETAAAKFE RQHMDSPDLG TLVPRGSMAD IGSEFMADER KDEAKAPHWT SAPLTEASAH
SHPPEIKDQG GAGEGLVRSA NGFPYREDEE GAFGEHGSQG TYSNTKENGI NGELTSADRE
TAEEVSARIV QVVTAEAVAV LKGEQEKEAQ HKDQTAALPL AAEETANLPP SPPPSPASEQ
TVTVEEAAGG ESALAPSVFK QAKDKVSDGV TKSPEKRSSL PRPSSILPPR RGVSGDRDEN
SFSLNSSISS SARRTTRSEP IRRAGKSGTS TPTTPGSTAI TPGTPPSYSS RTPGTPGTPS
YPRTPHTPGT PKSAILVPSE KKVAIIRTPP KSPATPKQLR LINQPLPDLK NVKSKIGSTD
NIKYQPKGGQ VQIVTKKIDL SHVTSKCGSL KNIRHRPGGG RVKIESVKLD FKEKAQAKVG
SLDNAHHVPG GGNVKIDSQK LNFREHAKAR VDHGAEIITQ SPGRSSVASP RRLSNVSSSG
SINLLESPQL ATLAEDVTAA LAKQGLVDKL AAALEHHHHHH

Number of amino acids: 521
Molecular weight: 55180.55
Theoretical pI: 9.23

Amino acid composition:
Ala (A) 52 10.0%
Arg (R) 29 5.6%
Asn (N) 16 3.1%
Asp (D) 22 4.2%
Cys (C) 1 0.2%
Gln (Q) 19 3.6%
Glu (E) 38 7.3%
Gly (G) 42 8.1%
His (H) 19 3.6%
Ile (I) 22 4.2%
Leu (L) 32 6.1%
Lys (K) 39 7.5%
Met (M) 4 0.8%
Phe (F) 8 1.5%
Pro (P) 46 8.8%
Ser (S) 59 11.3%
Thr (T) 37 7.1%
Trp (W) 1 0.2%
Tyr (Y) 5 1.0%
Val (V) 30 5.8%


Total number of negatively charged residues (Asp + Glu): 60
Total number of positively charged residues (Arg + Lys): 68

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

Ext. coefficient 12950
Abs 0.1% (=1 g/l) 0.235, assuming all pairs of Cys residues form cystines

Ext. coefficient 12950
Abs 0.1% (=1 g/l) 0.235, assuming all Cys residues are reduced

1. Dehmelt, H and Halpain, S. The MAP2/Tau family of microtubule-associated proteins.
Genome Biol. 204 (2005)

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