Heparan Sulfate Proteoglycan (HSPG)

Heparan sulfate proteoglycans (HSPGs) are found on cell surfaces and in the extracelluar matrix of all mammalian tissues. HS consists of an unbranched chain of repeating disaccharide units containing glucosamine and hexuronic acid. HS is structurally similar to heparin but contains lower contents of N-sulfates, O-sulfate, and IdUA, and more GlcUA than does heparin. HS is attached to serine residues at Ser-Gly sequences in a protein core through a tetrasaccharide linkage region. Generally two to four HS chains are attached to a protein core and CS may be found in some protein cores. HSPGs found in the brain are dystroglycan , N-syndecan glypican, and perlecan. The composition of HS isolated from brain differs significantly from that of other organs and is developmentally regulated. Thus, differences in protein cores, disaccharide composition, the extent and position of sulfation, and the number of GAG chains diversify chemical structure and functions of HSPGs. The postulated functions of HSPG include cell proliferation, differentiation, adhesion, migration, and morphogenesis.\n\nApplications:\nSuitable for use in ELISA, Western Blot and Immunohistochemistry. Not suitable for use on cultured cell. Other applications not tested. \n\nRecommended Dilutions: \nELISA: 1:50 \nWestern Blot: 1:50-1:100. Do not use milk as blocking agent. Must be digested with chondroitinase prior to running on SDS gels because undigested HSPG is too large for most gels. Treatment is at a concentration of chondroitinase of 10u/ml in Tris-HCl pH 8.0. Make tissue or cell extract in 20-50mM Tris pH 7.6-8.0 with 0.15M sodium chloride in the presence of protease inhibitors. Add 1ul of enzyme to 30ul of extract and incubate 30 minutes at 37C. Add SDS sample buffer, heat or boil sample as normal for SDS \nImmunohistochemistry: 1:25. Trypsin treatment required as antigen retrieval.\nOptimal dilutions to be determined by researcher. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

H1890-88

Size

1ml

Applications

ELISA, IHC, WB

Hosts

Mouse

Reactivities

Hum, Bov, Can, NHP

Form

Supplied as a liquid in DMEM, 3% BSA. No preservatives added.

P Type

Mab

Purity

Tissue culture supernatant

Isotype

IgG1

References

1. Dumon, KR, et al., Canc Research 61: 4827-4836 (2001). 2. Kemeny, E, et al., Connect Tissue Research 18(1):9-25. 3. Ortega-Barria, E, et al., J Biol Chem 274(3):1267-1276 (1999). 4. Wilkins, BS, et al., British Journal of Haematology 90:757

Additional Info

Recognizes bovine Heparan Sulfate Proteoglycan protein core of basement membrane at >200kD. Species Crossreactivity: human, canine and monkey. No crossreactivity with mouse, rat or amphibian tissues.