Anti-Heparan Sulfate Proteoglycan 2/Perlecan 抗体 [A7L6] (ab2501)

Reviews (3)Q&A (12)References (19)

製品の概要

  • 製品名

    Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6]
    Heparan Sulfate Proteoglycan 2/Perlecan 一次抗体 製品一覧

  • 製品の詳細

    Rat monoclonal [A7L6] to Heparan Sulfate Proteoglycan 2/Perlecan

  • 由来種

    Rat

  • 特異性

    Recognizes domain IV of heparan sulphate proteoglycan 2 / perlecan. The reactivity is independent of the galactosaminoglycan moieties. Therefore, the epitope is not sensitive to heparitinase treatment.

  • アプリケーション

    適用あり: ICC, IHC-P, IHC-Fr, IP, WBmore details

  • 種交差性

    交差種: Mouse, Rat, Cow, Human, Pig, Fish

  • 免疫原

    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.

  • 特記事項


    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.

     This product was previously labelled as Heparan Sulfate Proteoglycan 2

     

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能

    Integral 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.

  • 組織特異性

    Found in the basement membranes.

  • 関連疾患

    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.

  • 配列類似性

    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.

  • 翻訳後修飾

    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.

  • 細胞内局在

    Secreted > extracellular space > extracellular matrix > basement membrane.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Basement membrane specific heparan sulfate proteoglycan core protein antibody
    • Endorepellin (domain V region) antibody
    • Heparan sulfate proteoglycan of basement membrane antibody
    • HSPG 2 antibody
    • HSPG antibody
    • Hspg2 antibody
    • LG3 peptide antibody
    • Perlecan antibody
    • PGBM_HUMAN antibody
    • PLC antibody
    • Schwartz Jampel syndrome 1 (chondrodystrophic myotonia) antibody
    • SJA antibody
    • SJS antibody
    • SJS1 antibody
    see all

画像

  • Immunohistochemistry (Frozen sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6] (ab2501)
    Immunohistochemistry (Frozen sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6] (ab2501)

    IHC image on a frozen section of human colon showing strong reactivity in the extracellular matrix and basement membrane.

  • Immunohistochemistry (Frozen sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6] (ab2501)
    Immunohistochemistry (Frozen sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6] (ab2501)
    Immunohistochemistry on frozen section of Human kidney showing strong reactivity in the extracellular matrix and basement membrane.

参考文献

This product has been referenced in:

  • Horsley H  et al. A urine-dependent human urothelial organoid offers a potential alternative to rodent models of infection. Sci Rep 8:1238 (2018). Read more (PubMed: 29352171) »
  • Brauchle E  et al. Biomechanical and biomolecular characterization of extracellular matrix structures in human colon carcinomas. Matrix Biol 68-69:180-193 (2018). IHC ; Human . Read more (PubMed: 29605717) »
See all 19 Publications for this product

ab2501 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

レビューと Q&A

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1-10 of 15 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric buffer
Permeabilization
Yes - Triton 100
Specification
kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

投稿 Sep 18 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6]

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (heart and vascular tissue)
Permeabilization
No
Specification
heart and vascular tissue
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

投稿 Jul 01 2019

Immunohistochemistry (Resin sections) abreview for Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [A7L6]

 Average
Abreviews
abreview image
Application
Immunohistochemistry (Resin sections)
Sample
Human Tissue sections (Human tonsil)
Specification
Human tonsil
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Abcam user community

Verified customer

投稿 Dec 31 2013

Question

Phone calls reporting problems in using the anti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6](ab2501) in western blotting.

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Answer

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 with thisproducts. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As discussed over the phone, I have attached aquestionnaire so thatI can gather further information regarding the samples tested and the protocol used with this antibody. OnceI havereceive the completed questionnaire,I will look at the protocol and see if there are any suggestionsI can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. In order to understand the problem fully I would appreciate it if you could send any images you have collected with the antibody, and any other antibody you have used with the samples.

If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

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Question

Hallo,

Danke für ihre Antwort, bevor ich ihre ganzen Fragen beantworte hätte ich noch eine Frage, wäre es mgl das sie mir einen Marker für diese großen Proteine nennen? Ich konnte leider keine finden. Wir benutzen zum Transfer außerdem das System von Invitrogen das 7min dauert, mit Ponceau konnten wir zeigen das auch große Proeteine auf der Membran ware. Der Lysepuffer (1% Nonident P40, 500mmol/l Tris-Base (pH 7.6), 150mmol/lNaC). Zum Transferpuffer kann ich nicht soviel sagen da es keine gibt. Wir benutzen ja den iBlot ohne Puffer.

Ich hoffe sie können mir dennoch weiterhelfen,

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Answer

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

Da Sie ein sehr schweres Protein detektieren möchten, möchte ich Ihnen empfehlen, einen Ureapuffer zu verwenden. Ich habe Ihnen ein sehr interessantes Paper über die Löslichkeit von verschieden Proteinen hinsichtlich des Kompartiments und des Gewichtes an diese Email gehängt.

Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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Question

Hello,
I want to order the anti heparan sulfate antibody clone A7L6 (ab2501). But, I read on you data sheet that it is not sensitive to heparitinase treatment. My question is: is that antibody recogniez the heparan sulfate after degradation or cleavage by the heparanase ( an enzyme wich cleave the heparan sulfate in the 2-O-sulfate groupe on a hexuronic acid residue).
Best wishes

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Answer

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 literature (see attached), the antibody recognizes an epitope in domain IV which is stable after heparinase III (heparitinase I) treatment.

I am sorry that we don't have more information about this epitope, but if you have any further questions or if there is anything else that we can do for you, please don't hesitate to ask.

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Question

In detail, I would be interested in a BSA / gelatin free format of this antibody: “Anti-Heparan Sulphate Proteoglycan (Large) antibody [A7L6] (ab2501)” Please let me know if it is possible and if you would charge any additional costs. Best regards        

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Answer

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 following antibody purification kits: ab102784  Antibody Purification kit (3 purifications) Click here (or use the following: https://www.abcam.com/index.html?datasheet=102784).         ab102783  Antibody Purification kit (1 purification) Click here (or use the following: https://www.abcam.com/index.html?datasheet=102783). I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.    

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Question

>1) what tissue you used? human tonsils > >2) how long you fixed it for? Routine fixation performed by the pathology department from 4 hours to o/n in 4% formaldehyde >3) did you try the pretreatment with trypsin for >30 minutes at 37°C: 0.1% trypsin in 50mM >Tris-HCl + 0,1% CaCl2 pH 8.0? yes >4)how long did you incubate the antibody for? 1h30 at RT >5)what dilution buffer did you use? PBS 1% BSA >6)did you perform amplification with an ABC kit? yes >7) If so did you quench endogenous peroxidases yes >8)did you perform a no primary control? yes

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Answer

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 incubation I would be prepared in this instance to offer you a refund/credit note even though the Abpromise of 90 days covering the antibody has expired, according to our records. I look forward to hearing how you get on with an overnight incubation,

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Question

What is recommended as a positive control for Western blotting? What type of gel should be used? What size band is detected?

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Answer

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).

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Question

I have a question regarding the antibody ab2501 anti-heparan sulphate Proteoglycan. We wanted to use the antibody for Paraffine staining we have it in house now. Can you tell me which antigen retrieval is the best one to use. Or if its already done in an artikel can you sent me a link to the artikel.

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Answer

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 need anything further or any help then please let us know.

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