Ordering Information
- : MAB458
- : 1 mL
- : 100 μg/mL
Anti-Heparan Sulfate Proteoglycan, clone 7E12
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| B, Ca, H, Mk, not Am, not M, not R | ELISA, IH | Mouse | Culture Supernatant | Monoclonal Antibody |
Description:
Anti-Heparan Sulfate Proteoglycan, clone 7E12
Trade Name:
Chemicon (Millipore)
Specificity:
Heparan Sulfate Proteoglycan protein core of basement membrane.
Tátrai et al. (2006) have demonstrated that this antibody also shows specificity to agrin.
SPECIES REACTIVITIES:
No cross-reactivity with mouse, rat or amphibian tissues.
Tátrai et al. (2006) have demonstrated that this antibody also shows specificity to agrin.
SPECIES REACTIVITIES:
No cross-reactivity with mouse, rat or amphibian tissues.
Immunogen:
Bovine Glomeruli Heparan Sulfate Proteoglycan.
Clone:
7E12
Isotype:
IgG1
Background Information:
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. Thus 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. HSPGs differ in their protein cores that are thought to determine the location of HSPG in the cell membrane (syndecan and glypican) and extracellular matrix (perlecan and dystroglycan). 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.
Species Reactivity:
- Bovine
- Canine
- Human
- Monkey
Non-Reactive Species:
- Amphibian
- Mouse
- Rat
Application Notes:
ELISA at 1:50
Western Blot at 1:50-1:100. Do not place antibody in milk block as absorbtion can occur, also samples 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 NaCl in the presence of protease inhibitors. Add 1 microliter of enzyme to 30 microliters of extract and incubate 30 minutes at 37C. Then add SDS sample buffer, heat or boil sample as normal for SDS reducing samples.
Antibody detects >200 kDa core protein, typically as a large smear.
Immunohistochemistry at 1:25: with trypsin antigen recovery, recommended. Heparitinase appears to enhance 7E12 immunoreactivity as well.
Not suggested for use on cultured cells or culture extracts, only in tissue. Working dilutions are suggested only and are for use in conjunction with purified HSPG.
Optimal working dilutions must be determined by end user.
Western Blot at 1:50-1:100. Do not place antibody in milk block as absorbtion can occur, also samples 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 NaCl in the presence of protease inhibitors. Add 1 microliter of enzyme to 30 microliters of extract and incubate 30 minutes at 37C. Then add SDS sample buffer, heat or boil sample as normal for SDS reducing samples.
Antibody detects >200 kDa core protein, typically as a large smear.
Immunohistochemistry at 1:25: with trypsin antigen recovery, recommended. Heparitinase appears to enhance 7E12 immunoreactivity as well.
Not suggested for use on cultured cells or culture extracts, only in tissue. Working dilutions are suggested only and are for use in conjunction with purified HSPG.
Optimal working dilutions must be determined by end user.
Presentation:
Liquid tissue culture supernatant, containing 3% BSA and DMEM media base with no preservatives.
Storage Conditions:
Maintain at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze/thaw cycles.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- HSPG2
- SJA
- perlecan
- SJS
- HSPG
- PRCAN
- Perlecan
- SJS1
- PLC
Alternate Names:
HSPG
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
- ELISA
- Immunohistochemistry
Entrez Gene Summary:
Heparan sulfate proteoglycan is a major component of basement membranes, where the molecule may be involved in the stabilization of other molecules as well as being involved with glomerular permeability to macromolecules and cell adhesion. This form of HSPG, known as HSPG2 or perlecan, is encoded by a gene that maps to chromosome 1. The gene for the form of HSPG associated with the cell surface of fibroblasts has been mapped to human chromosome 8 (MIM 142460).[supplied by OMIM]
UniProt Summary:
FUNCTION: SwissProt: P98160 # This protein is an integral component of basement membranes. It is responsible for the fixed negative electrostatic charge and is involved in the charge-selective ultrafiltration properties. It serves as an attachment substrate for cells.
SIZE: 4391 amino acids; 468825 Da
SUBUNIT: Purified perlecan has a strong tendency to aggregate in dimers or stellate structures. It interacts with other basement membrane components such as laminin, prolargin and collagen type IV. Interacts with COL13A1, FGFBP1 and VWA1.
SUBCELLULAR LOCATION: Secreted, extracellular space.
TISSUE SPECIFICITY: Found in the basement membranes.
PTM: N- and O-glycosylated; contains three heparan sulfate chains.
DISEASE: SwissProt: P98160 # Defects in HSPG2 are the cause of Schwartz-Jampel syndrome (SJS1) [MIM:255800]; a rare autosomal recessive disorder characterized by permanent myotonia (prolonged failure of muscle relaxation) and skeletal dysplasia, resulting in reduced stature, kyphoscoliosis, bowing of the diaphyses and irregular epiphyses. & Defects in HSPG2 are the cause of dyssegmental dysplasia Silverman-Handmaker type (DDSH) [MIM:224410]. The dyssegmental dysplasias are rare, autosomal recessive skeletal dysplasias with anisospondyly and micromelia. There are two recognized types: the severe, lethal DDSH and the milder Rolland-Desbuquois form. Individuals with DDSH also have a flat face, micrognathia, cleft palate and reduced joint mobility, and frequently have an encephalocoele. The endochondral growth plate is short, the calcospherites (which are spherical calcium-phosphorus crystals produced by hypertrophic chondrocytes) are unfused, and there is mucoid degeneration of the resting cartilage.
SIMILARITY: SwissProt: P98160 ## Contains 4 EGF-like domains. & Contains 22 Ig-like C2-type (immunoglobulin-like) domains. & Contains 11 laminin EGF-like domains. & Contains 3 laminin G-like domains. & Contains 3 laminin IV type A domains. & Contains 4 LDL-receptor class A domains. & Contains 1 SEA domain.
SIZE: 4391 amino acids; 468825 Da
SUBUNIT: Purified perlecan has a strong tendency to aggregate in dimers or stellate structures. It interacts with other basement membrane components such as laminin, prolargin and collagen type IV. Interacts with COL13A1, FGFBP1 and VWA1.
SUBCELLULAR LOCATION: Secreted, extracellular space.
TISSUE SPECIFICITY: Found in the basement membranes.
PTM: N- and O-glycosylated; contains three heparan sulfate chains.
DISEASE: SwissProt: P98160 # Defects in HSPG2 are the cause of Schwartz-Jampel syndrome (SJS1) [MIM:255800]; a rare autosomal recessive disorder characterized by permanent myotonia (prolonged failure of muscle relaxation) and skeletal dysplasia, resulting in reduced stature, kyphoscoliosis, bowing of the diaphyses and irregular epiphyses. & Defects in HSPG2 are the cause of dyssegmental dysplasia Silverman-Handmaker type (DDSH) [MIM:224410]. The dyssegmental dysplasias are rare, autosomal recessive skeletal dysplasias with anisospondyly and micromelia. There are two recognized types: the severe, lethal DDSH and the milder Rolland-Desbuquois form. Individuals with DDSH also have a flat face, micrognathia, cleft palate and reduced joint mobility, and frequently have an encephalocoele. The endochondral growth plate is short, the calcospherites (which are spherical calcium-phosphorus crystals produced by hypertrophic chondrocytes) are unfused, and there is mucoid degeneration of the resting cartilage.
SIMILARITY: SwissProt: P98160 ## Contains 4 EGF-like domains. & Contains 22 Ig-like C2-type (immunoglobulin-like) domains. & Contains 11 laminin EGF-like domains. & Contains 3 laminin G-like domains. & Contains 3 laminin IV type A domains. & Contains 4 LDL-receptor class A domains. & Contains 1 SEA domain.
Brand Family:
Chemicon
Protein/Isoform Description:
Heparan Sulfate is a glycosaminoglycan, as is heparin. These proteoglycans bind proteins, including P-Selectin, and participate in the internalization of lipoproteins. It is thought that the protein-proteoglycan binding affinity comes from ionic interactions and the entropy
generated by the release of cations upon
protein binding
generated by the release of cations upon
protein binding
Product Name:
Anti-Heparan Sulfate Proteoglycan, clone 7E12
Concentration:
100 μg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
1 mL
Format:
Culture Supernatant
Host:
Mouse