Recombinant Human COMP protein, FLAG-tagged

• Cat.No.: COMP-3659H
• Product Overview: Recombinant Human COMP fused with FLAG tag at N-terminal was expressed in HEK293.

Specification

• Species: Human
• Source: HEK293
• Tag: Flag
• Protein Length: 750 AA
• Description: The protein encoded by this gene is a noncollagenous extracellular matrix (ECM) protein. It consists of five identical glycoprotein subunits, each with EGF-like and calcium-binding (thrombospondin-like) domains. Oligomerization results from formation of a five-stranded coiled coil and disulfides. Binding to other ECM proteins such as collagen appears to depend on divalent cations. Mutations can cause the osteochondrodysplasias pseudochondroplasia (PSACH) and multiple epiphyseal dysplasia (MED).
• Form: Filtered (0.4 μm) and lyophilized from 0.5 mg/ml in 20mM Tris buffer, 50mM NaCl, pH 7.5
• Molecular Mass: 82.4 kDa (calculated)
• AA Sequence: HVDYKDDDDK PAGQGQSPLG SDLGPQMLRE LQETNAALQD VRELLRQQVR EITFLKNTVM ECDACGMQQS VRTGLPSVRP LLHCAPGFCF PGVACIQTES GARCGPCPAG FTGNGSHCTD VNECNAHPCF PRVRCINTSP GFRCEACPPG YSGPTHQGVG LAFAKANKQV CTDINECETG QHNCVPNSVC INTRGSFQCG PCQPGFVGDQ ASGCQRRAQR FCPDGSPSEC HEHADCVLER DGSRSCVCAV GWAGNGILCG RDTDLDGFPD EKLRCPERQC RKDNCVTVPN SGQEDVDRDG IGDACDPDAD GDGVPNEKDN CPLVRNPDQR NTDEDKWGDA CDNCRSQKND DQKDTDQDGR GDACDDDIDG DRIRNQADNC PRVPNSDQKD SDGDGIGDAC DNCPQKSNPD QADVDHDFVG DACDSDQDQD GDGHQDSRDN CPTVPNSAQE DSDHDGQGDA CDDDDDNDGV PDSRDNCRLV PNPGQEDADR DGVGDVCQDD FDADKVVDKI DVCPENAEVT LTDFRAFQTV VLDPEGDAQI DPNWVVLNQG REIVQTMNSD PGLAVGYTAF NGVDFEGTFH VNTVTDDDYA GFIFGYQDSS SFYVVMWKQM EQTYWQANPF RAVAEPGIQL KAVKSSTGPG EQLRNALWHT GDTESQVRLL WKDPRNVGWK DKKSYRWFLQ HRPQVGYIRV RFYEGPELVA DSNVVLDTTM RGGRLGVFCF SQENIIWANL RYRCNDTIPE DYETHQLRQA
• Endotoxin: < 0.1 EU/ug
• Purity: Purity as determined by densitometric image analysis: >90%
• Applications: Cell culture and/or animal studies, ELISA, Western blotting
• Storage: Store lyophilized protein at –80 centigrade. Lyophilized protein remains stable until the expiry date when stored at –80 centigrade. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at –80 centigrade for long term storage. Reconstituted protein can be stored at 4 centigrade for a limited period of time; it does not show any change after one week at 4 centigrade.
• Reconstitution: Add deionized water to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. Filter sterilize your culture media/working solutions containing this non-sterile product before using in cell culture.
• Publications:
  Secretory cargo sorting by Ca 2+-dependent Cab45 oligomerization at the trans-Golgi network. (2016)

Gene Information

• Gene Name: COMP cartilage oligomeric matrix protein [ Homo sapiens ]
• Official Symbol: COMP
• Synonyms: COMP; cartilage oligomeric matrix protein; cartilage oligomeric matrix protein (pseudoachondroplasia, epiphyseal dysplasia 1, multiple) , EDM1, EPD1, PSACH; MED; THBS5; thrombospondin 5; TSP5; thrombospondin-5; pseudoachondroplasia (epiphyseal dysplasia 1, multiple); cartilage oligomeric matrix protein(pseudoachondroplasia, epiphyseal dysplasia 1, multiple); cartilage oligomeric matrix protein (pseudoachondroplasia, epiphyseal dysplasia 1, multiple); EDM1; EPD1; PSACH; MGC131819; MGC149768
• Gene ID: 1311
• mRNA Refseq: NM_000095
• Protein Refseq: NP_000086
• MIM: 600310
• UniProt ID: P49747
• Chromosome Location: 19p13.1
• Pathway: ECM-receptor interaction, organism-specific biosystem; ECM-receptor interaction, conserved biosystem; Focal Adhesion, organism-specific biosystem; Focal adhesion, organism-specific biosystem; Focal adhesion, conserved biosystem; Malaria, organism-specific biosystem; Malaria, conserved biosystem
• Function: calcium ion binding; collagen binding; extracellular matrix structural constituent; heparan sulfate proteoglycan binding; heparin binding; protein binding

Citation

Secretory cargo sorting by Ca 2+ -dependent Cab45 oligomerization at the trans-Golgi network

Journal: The Journal of Cell Biology    PubMed ID: 27138253    Data: 2016/5/9

Authors: Alvaro H. Crevenna, Birgit Blank, Julia von Blume

Article Snippet:For STORM measurements Cy5-conjugated AffiniPure (rabbit, 711–175-152; Jackson ImmunoResearch Laboratories, Inc.) was used for Cab45 as a secondary antibody and Alexa Fluor 488 sheep (A11015) for TGN46 (Thermo Fisher Scientific).For STORM measurements Cy5-conjugated AffiniPure (rabbit, 711–175-152; Jackson ImmunoResearch Laboratories, Inc.) was used for Cab45 as a secondary antibody and Alexa Fluor 488 sheep (A11015) for TGN46 (Thermo Fisher Scientific).. The recombinant proteins COMP-FLAG (COMP-3659H; Creative BioMart), COMP (ab104358; Abcam), LysozymeC (Sigma-Aldrich), and LysozymeC-Cy5 (LS1-S5-1; Nanocs) were used for in vitro analysis.

Cab45 Ca 2+ binding is required for sorting in living cells. (A) Cell lysates of Cab45 ?/? , Cab45 ?/? Cab45-HA WT or Cab45 ?/? Cab45-HA 6EQ-mut HeLa cells were analyzed by WB with a Cab45 antibody. (B and D) Cell culture supernatants of the same cells were incubated in serum-free medium for 4 h at 37°C. Media and cell extracts were blotted and probed with antibodies against COMP (B) or Flag (LyzC; D). Western blots from three independent experiments were quantified by densitometry using the ImageJ software. Bar graphs represent the densitometry values of external COMP (C) and LyzC (E) normalized to internal protein contents, respectively. Error bars represent SD calculated of three independent experiments of secreted COMP (C) or LyzC (E), respectively ( n = 3). **, P < 0.01 (compared datasets).