Anti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6] (ab2501)

Overview

  • Product nameAnti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6]
    See all Heparan Sulphate Proteoglycan (Large) primary antibodies
  • Description
    Rat monoclonal [A7L6] to Heparan Sulphate Proteoglycan (Large)
  • SpecificityRecognizes domain IV of the core protein of the large heparan sulphate proteoglycan or perlecan. The reactivity is independent of the galactosaminoglycan moieties. Therefore, the epitope is not sensitive to heparitinase treatment.
  • Tested applicationsSuitable for: ICC, IHC-P, IHC-Fr, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Pig, Fish
  • Immunogen

    High molecular mass material derived from the Engelbreth-Holm-Swarm (EHS) tumor matrix containing laminin, entactin and HSPG. The antibody was raised against the total Heparan Sulphate Proteoglycan protein; no epitope mapping was ever performed.

  • General notes


    Proteoglycans are macromolecules consisting of a variety of core proteins with covalently attached one or several polysaccharide chains of the glycosaminoglycan type (heparan sulphate, heparin, chondroitin sulphate, dermatan sulphate or keratan sulphate). At least two forms of basement membrane heparan sulphate proteoglycan (HSPG) have been identified. One with a large core protein (> 400 kD) and one with a small core protein (30 kD). The large HSPG is probably the most abundant basement membrane proteoglycan. It is located predominantly in the lamina lucida, where it forms clustered aggregates and interacts with other basement membrane components to form the matrix. In addition, it also plays a critical role in attachment of cells to the basal membrane via integrin receptors.

Properties

Applications

Our Abpromise guarantee covers the use of ab2501 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC 1/25 - 1/200. Use amplification with ABC (avidin biotin complex).
IHC-P Use at an assay dependent dilution. Perform enzymatic antigen retrieval before commencing with IHC staining protocol. Use amplification with ABC (avidin biotin complex).
IHC-Fr Use at an assay dependent dilution. PubMed: 16055495Use amplification with ABC (avidin biotin complex).
IP Use at an assay dependent dilution.
WB 1/100 - 1/1000.

Target

  • FunctionIntegral component of basement membranes. Component of the glomerular basement membrane (GBM), responsible for the fixed negative electrostatic membrane charge, and which provides a barrier which is both size- and charge-selective. It serves as an attachment substrate for cells. Plays essential roles in vascularization. Critical for normal heart development and for regulating the vascular response to injury. Also required for avascular cartilage development.
    Endorepellin in an anti-angiogenic and anti-tumor peptide that inhibits endothelial cell migration, collagen-induced endothelial tube morphogenesis and blood vessel growth in the chorioallantoic membrane. Blocks endothelial cell adhesion to fibronectin and type I collagen. Anti-tumor agent in neovascularization. Interaction with its ligand, integrin alpha2/beta1, is required for the anti-angiogenic properties. Evokes a reduction in phosphorylation of receptor tyrosine kinases via alpha2/beta1 integrin-mediated activation of the tyrosine phosphatase, PTPN6.
    The LG3 peptide has anti-angiogenic properties that require binding of calcium ions for full activity.
  • Tissue specificityFound in the basement membranes.
  • Involvement in diseaseDefects 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.
  • Sequence similaritiesContains 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.
  • Post-translational
    modifications
    Proteolytic processing produces the C-terminal angiogenic peptide, endorepellin. This peptide can be further processed to produce the LG3 peptide.
    N- and O-glycosylated; contains three heparan sulfate chains. The LG3 peptide contains at least three and up to five potential O-glycosylation sites but no N-glycosylation.
  • Cellular localizationSecreted > extracellular space > extracellular matrix > basement membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Heparan sulfate proteoglycan of basement membrane antibody
    • HSPG antibody
    • Hspg2 antibody
    • LG3 peptide antibody
    • Perlecan antibody
    • PGBM_HUMAN antibody
    • PLC antibody
    • SJS1 antibody
    see all

Anti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6] images

  • ab2501 staining Heparan Sulphate Proteoglycan (Large) in human tonsil tissue sections by Immunohistochemistry (resin sections). Samples were incubated with primary antibody (1/100 in TBS) for 16 hours at 25°C. A biotin-conjugated rabbit anti-rat IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunohistochemistry on frozen section of Human kidney showing strong reactivity in the extracellular matrix and basement membrane.

References for Anti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6] (ab2501)

This product has been referenced in:
  • Tian C  et al. A splice mutation and mRNA decay of EXT2 provoke hereditary multiple exostoses. PLoS One 9:e94848 (2014). WB ; Human . Read more (PubMed: 24728384) »
  • Saxena A  et al. CXCR3-independent actions of the CXC chemokine CXCL10 in the infarcted myocardium and in isolated cardiac fibroblasts are mediated through proteoglycans. Cardiovasc Res 103:217-27 (2014). Mouse . Read more (PubMed: 24891401) »

See all 12 Publications for this product

Product Wall

Application Immunohistochemistry (Resin sections)
Sample Human Tissue sections (Human tonsil)
Specification Human tonsil
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Abcam user community

Verified customer

Submitted Dec 31 2013

Thank you for calling us yesterday and reporting the problems you have encountered in using the anti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6](ab2501) in western blotting.

I am sorry to hear youhave beenexperiencing difficulties ...

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Vielen Dank für Ihre Anfrage.

Leider bieten wir keinen Marker an, der höher als 225 kDa läuft. Allerdings habe ich einen Marker bis 300kDa bei Fermentas gefunden:

http://www.fermentas.de/product_info.php?info=p1496&language=de
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Thank you for contacting us with your question and for your patience while I have looked into your enquiry.

Unfortunatelywe don't have any information regarding reactivity of this clone following herapanase treatment. According to the literat...

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Thank you for your enquiry. I am sorry to confirm that we are not able to provide this particular antibody BSA and glycerol free. I can suggest you may like to consider purifying the antibody in order to remove the BSA and glycerol. We do have the ...

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Thank you for your reply and providing more protocol information. I would recommend to incubate the antibody overnight at 4C as this will promote the antibody binding to the protein of interrest. If you still experience problems with the overnight i...

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Best to use lysate of mouse EHS tumor cells as cultured cells do not produce much of Heparan sulphate proteoglycan. Try to use 7.5% gel as the molecule is very large (the large core protein is 400 kDa).

Thank you for your enquiry. We would suggest applying protease digestion. For details on IHC and antigen retrieval, please click on the Resources section on the top panel of the Abcam homepage, you might find some useful information there. If you...

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Thank you for forwarding the complaint on-line. I have read through the protocol which seems to be absolutely fine. Looking at the order record, we have sold several vials of this batch without any problem. The antibody should recognize Heparan Sul...

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Thank you for your enquiry. All the information that we have regarding this antibody is located on the online datasheet. Formalin fixed tissue requires an antigen retrieval step before immunohistochemical staining can proceed; if no antigen retrieval s...

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