製品の概要

  • 製品名
    Anti-Elastin antibody [BA-4]
    Elastin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [BA-4] to Elastin
  • 由来種
    Mouse
  • アプリケーション
    適用あり: WB, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Sheep, Goat, Guinea pig, Cat, Dog, Human, Pig
  • 免疫原

    Bovine alpha-Elastin.

  • ポジティブ・コントロール
    • WB: Human skin tissue lysate. IHC-P: FFPE normal Human Duodenum tissue sections IHC-Fr: Human normal skin
  • 特記事項

     

    Elastin is an important polymeric protein of connective tissue that imparts elasticity to vertebrate elastic tissues.

     

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab9519 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
WB Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 68 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr Use a concentration of 1 µg/ml.

ターゲット情報

  • 機能
    Major structural protein of tissues such as aorta and nuchal ligament, which must expand rapidly and recover completely. Molecular determinant of the late arterial morphogenesis, stabilizing arterial structure by regulating proliferation and organization of vascular smooth muscle.
  • 組織特異性
    Expressed within the outer myometrial smooth muscle and throughout the arteriolar tree of uterus (at protein level). Also expressed in the large arteries, lung and skin.
  • 関連疾患
    Defects in ELN are a cause of autosomal dominant cutis laxa (ADCL) [MIM:123700]. Cutis laxa is a rare connective tissue disorder characterized by loose, hyperextensible skin with decreased resilience and elasticity leading to a premature aged appearance. The skin changes are often accompanied by extracutaneous manifestations, including pulmonary emphysema, bladder diverticula, pulmonary artery stenosis and pyloric stenosis.
    Defects in ELN are the cause of supravalvular aortic stenosis (SVAS) [MIM:185500]. SVAS is a congenital narrowing of the ascending aorta which can occur sporadically, as an autosomal dominant condition, or as one component of Williams-Beuren syndrome.
    Note=ELN is located in the Williams-Beuren syndrome (WBS) critical region. WBS results from a hemizygous deletion of several genes on chromosome 7q11.23, thought to arise as a consequence of unequal crossing over between highly homologous low-copy repeat sequences flanking the deleted region. Haploinsufficiency of ELN may be the cause of certain cardiovascular and musculo-skeletal abnormalities observed in the disease.
  • 配列類似性
    Belongs to the elastin family.
  • 翻訳後修飾
    Elastin is formed through the cross-linking of its soluble precursor tropoelastin. Cross-linking is initiated through the action of lysyl oxidase on exposed lysines to form allysine. Subsequent spontaneous condensation reactions with other allysine or unmodified lysine residues result in various bi-, tri-, and tetrafunctional cross-links. The most abundant cross-links in mature elastin fibers are lysinonorleucine, allysine aldol, desmosine, and isodesmosine.
    Hydroxylation on proline residues within the sequence motif, GXPG, is most likely 4-hydroxy as this fits the requirement for 4-hydroxylation in vertebrates.
  • 細胞内局在
    Secreted > extracellular space > extracellular matrix. Extracellular matrix of elastic fibers.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Elastin antibody
    • ELN antibody
    • ELN_HUMAN antibody
    • SVAS antibody
    • Tropoelastin antibody
    • WBS antibody
    • WS antibody
    see all

画像

  • IHC image of Elastin staining in a section of formalin-fixed paraffin-embedded normal human normal duodenum* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab9519, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • IHC image of ab9519 staining in 10% formaldehyde fixed frozen tissue section of normal human skin.

    Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with ab9519 (1μg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647)) and DAPI for 1 hour at room temperature.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab9519. 

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

  • ab9519 staining pig small intestine tissue sections by IHC-P.  Sections were fixed with Bouins and subjected to an enzymatic antigen retreival step.  The primary antibody was diluted 1/200 in a commercially available antibody diluent (replacing the blocking step) and incubated with the sample for 1 hour at 20°C.  A HRP conjugated goat anti-mouse antibody was used as the secondary.

    See Abreview

  • Anti-Elastin antibody [BA-4] (ab9519) at 5 µg/ml + Human skin tissue lysate - total protein (ab30166) at 20 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes

参考文献

This product has been referenced in:
  • Godinho MSC  et al. Elastin is Localised to the Interfascicular Matrix of Energy Storing Tendons and Becomes Increasingly Disorganised With Ageing. Sci Rep 7:9713 (2017). Read more (PubMed: 28855560) »
  • Kawamoto-Hirano A  et al. Cricoarytenoid Articulation in Elderly Japanese With Special Reference to Morphology of the Synovial Tissue. Ann Otol Rhinol Laryngol 125:219-27 (2016). Read more (PubMed: 26391093) »
See all 12 Publications for this product

レビューと Q&A

1-10 of 14 Q&A

Question
Answer

Thank you for your enquiry.

I can confirm that ab21640 and ab9519 are sold as whole antiseruma and ascites respectively. Unpurified antibodies, such as these sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for whole antiserum, concentration of antibody is known to very between 1 - 10 mg/ml, while the Anti-Elastin antibody [BA-4] (ab9519) is estimated to be around 0.1-0.2 mg/mL.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Question
Answer

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab9519 in rat. This code will give you: 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for ratand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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Question
Answer

I am very pleased to hear you would like to accept our offer and test ab9519 in mouse. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview formouse and include this code in the Additional Comments section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question
Answer

Vielen Dank für Ihren Anruf.


Ich habe einen Proteinsequenz Vergleich zwischen den Immunogenen derMaus Klone [BA-4] und[10B8] (ab9519, ab97963und ab77804)durchgeführt, und folgendes hat sich ergeben: Die Übereinstimmung zwischen dem Immunogen und demRatten Protein beträgt höchstens%. Damit ein Antikörper kreuzreagiert, sollte die Übereinstimmung mindestens 85% betragen. Es daher unwahrscheinlich, dass diese Antikörper IhrZielproteindetektieren werden.

Allerdings haben wir einen polyklonalenKaninchen Antikörper, der sowohl an der Ratte als auch in der ICC getestet wurde und dazu noch eine hohe Spezifität aufweist.

https://www.abcam.com/index.html?datasheet=23748 (or use the following: https://www.abcam.com/index.html?datasheet=23748).


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

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Question
Answer

Merci de votre appel et de votre intérêt pour nos anticorps contre l'élastine. Comme discuté au téléphone, nous n'avons malheureusement pas d’échantillon gratuit à des fins de tests préliminaires. Cependant, si vous souhaitez tester un de nos anticorps contre l'élastine sur le poisson, il me sera possible de vous offrir une remise. Après soumission d’une Abreview avec vos résultats, une remise correspondant à la valeur d’achat de l'anticorps acheté pourra être utilisable lors d’une prochaine commande d’anticorps primaire. Nous avons par exemple le ab9519, anticorps monoclonal de souris, testé en coupes paraffine, et sur les espèces suivantes: Hu, Cat, Dog, Goat, Gpig, Pig, Shp. Click here (or use the following: https://www.abcam.com/index.html?datasheet=9519) ou l'anticorps polyclonal, le ab21610 testé aussi en coupes paraffine et sur les éspèces suivantes  ö Hu, Rat, Chk, Cow, Dog, Pig Click here (or use the following: https://www.abcam.com/index.html?datasheet=21610). Étapes à suivre pour participer à cette offre : 1. Nous confirmer quel anticorps vous souhaiteriez tester chez le poisson afin de recevoir le code de remise correspondant. Ce code doit être édité avant l’achat de cet anticorps contre l'élastine. 2. Acheter l'anticorps contre l'élastine par téléphone, fax ou internet (www.abcam.com) 3. Le tester chez le poisson 4. Nous faire part de vos résultats, positifs ou négatifs, grâce à notre système Abreview. Pour plus d’informations https://www.abcam.com/abreviews 5. Après soumission de votre Abreview, appeler notre service clientèle afin de passer votre prochaine commande grâce au code de réduction. Ce code de réduction est utilisable 120 jours après son édition et utilisable lors de l’achat d’un autre anticorps primaire. Même si l'anticorps contre l'élastine est démontré ne pas fonctionner chez le poisson, après soumission de votre Abreview, vous recevrez quand même une remise sur l’achat de votre prochain anticorps. N’hésitez pas à nous demander plus d’informations concernant cette offre promotionnelle. Termes et Conditions : www.abcam.com/collaborationdiscount.  

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. With this specific antibody it has been shown that the method of antigen retrieval can be crucial in its performance and enzymatic retrieval is the most efficient. However, having reviewed the protocol details, I believe this product should have given satisfactory results with the methods used and unfortunately cannot find any additional suggestions to make. May I ask, did the positive control you carried out with the pericardium also give a negative result? It appears that you may have received a faulty vial of this antibody. I apologise for the inconvenience caused by this and would like to offer you a free of charge replacement of this antibody with a different lot or if this is unsatisfactory I can offer a credit note. In order to process this can you please let me know the original order number, the date on which it was placed or the address used to process it. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Answer

Thanks for your reply. I'm sorry but the specific enzyme used for antigen retrieval with ab9519 is not known. I think it would be useful to try pepsin or trypsin as well. Also, what type of sample tissue are you working with?

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Answer

Thank you for your enquiry. I just want to make sure I understand your question. You are wondering what enzyme should be conjugated to your secondary antibody for use in IHC-P? If that is the case, most commonly researchers use HRP. For use with the primary antibody ab9519, you could use ab98799, an HRP-conjugated secondary antibody. I hope this is helpful. Please contact me again if you have any further questions.

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Answer

Thank you for your enquiry. I have checked with the originator of the product but unfortunately, we are not sure about the isoforms detected by this antibody. However, I have managed to locate a publication on the subject and have listed it below for your reference. David et al. Characterization of Biologically Active Domains on Elastin: Identification of a Monoclonal Antibody to a Cell Recognition Site. Biochemistry 1986, 25, 5 172-5 176 http://pubs.acs.org/cgi-bin/abstract.cgi/bichaw/1986/25/i18/f-pdf/f_bi00366a028.pdf I hope the above information will be useful to you. Should you need any further assistance, please do not hesitate to contact me.

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Question
Answer

No high temperature antigen retrieval but proteolytic treatment with pepsin or trypsin should be OK. Please contact us again if you have any additional questions.

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1-10 of 14 Q&A

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