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    products/primary-antibodies/biotin-collagen-iv-antibody-ab6581.pdf

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Signal Transduction Cytoskeleton / ECM Basal Lamina
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Biotin Anti-Collagen IV antibody (ab6581)

  • Datasheet
  • SDS
Reviews (1)Q&A (6)References (7)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Collagen IV antibody (ab6581)

    Key features and details

    • Biotin Rabbit polyclonal to Collagen IV
    • Suitable for: ELISA, WB, IP, IHC-P
    • Reacts with: Cow, Human
    • Conjugation: Biotin
    • Isotype: IgG

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    Overview

    • Product name

      Biotin Anti-Collagen IV antibody
      See all Collagen IV primary antibodies
    • Description

      Biotin Rabbit polyclonal to Collagen IV
    • Host species

      Rabbit
    • Conjugation

      Biotin
    • Specificity

      negligible cross-reactivity with Type I, II, III, V or VI collagens. Non-specific cross reaction of anti-collagen antibodies with other human serum proteins or non-collagen extracellular matrix proteins is negligible.
    • Tested applications

      Suitable for: ELISA, WB, IP, IHC-Pmore details
    • Species reactivity

      Reacts with: Cow, Human
      Predicted to work with: Mammals
    • Immunogen

      Full length native protein (purified) corresponding to Collagen IV. Collagen Type IV from human and bovine placenta.

    • General notes

      At least 11 genetically distinct gene products are collectively referred to as 'collagen types' or other proteins and proteoglycans of the extracellular matrix. In humans, collagens are composed of about 20 unique protein chains which under go various types of post-translational modifications and are ultimately assembled into a triple helix. This results in great diversity between collagen types. Collagens are highly conserved throughout evolution and are characterized by an uninterrupted "Glycine-X-Y" triplet repeat that is a necessary part of the triple helical structure. For these reasons it is often extremely difficult to generate antibodies with specificities to collagens. The development of type specific antibodies is dependent on NON-DENATURED three-dimensional epitopes. This preparation results in a native conformation of the protein.


      These antibodies are well suited to detect extracellular matrix proteins in normal as well as disease state tissues. Disruption of tissue organization is the hallmark of neoplasia. Malignant lesions can be distinguished from benign by examining the breakdown of basement membranes and loss of 3-dimensional architecture. Malignant cells are presumed to use matrix metalloproteases to degrade barriers created by the extracellular matrix which then allows metastasis to occur. Collagenases, stomelysins and gelatinases can collectively degrade all of the various components of the extracellular matrix, including fibrillar and non-fibrillar collagens and basement membrane glycoproteins.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
    • Storage buffer

      Preservative: 0.01% Sodium azide
      Constituents: 0.44% Sodium chloride, 1% BSA, 4.77% Sodium borate, 0.146% EDTA
    • Concentration information loading...
    • Purity

      Immunogen affinity purified
    • Purification notes

      Immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against other collagens, human serum proteins and non-collagen extracellular matrix proteins to remove any unwanted specificities.
    • Primary antibody notes

      These antibodies are well suited to detect extracellular matrix proteins in normal as well as disease state tissues. Disruption of tissue organization is the hallmark of neoplasia. Malignant lesions can be distinguished from benign by examining the breakdown of basement membranes and loss of 3-dimensional architecture. Malignant cells are presumed to use matrix metalloproteases to degrade barriers created by the extracellular matrix which then allows metastasis to occur. Collagenases, stomelysins and gelatinases can collectively degrade all of the various components of the extracellular matrix, including fibrillar and non-fibrillar collagens and basement membrane glycoproteins.
    • Clonality

      Polyclonal
    • Isotype

      IgG
    • Research areas

      • Signal Transduction
      • Cytoskeleton / ECM
      • Basal Lamina
      • Signal Transduction
      • Cytoskeleton / ECM
      • Extracellular Matrix
      • ECM Proteins
      • Collagen
      • Stem Cells
      • Mesenchymal Stem Cells
      • Chondrogenesis
      • Cancer
      • Invasion/microenvironment
      • ECM
      • Extracellular matrix
      • Other

    Associated products

    • Isotype control

      • Biotin Rabbit IgG - Isotype Control (ab200208)
    • Positive Controls

      • Native Cow Collagen IV protein (ab7529)
      • Native Human Collagen IV protein (ab7536)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab6581 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
    ELISA (1)
    Use at an assay dependent concentration.
    WB
    Use at an assay dependent concentration. Predicted molecular weight: 161 kDa.

    Not recommended for use under denaturing conditions.

    IP
    Use at an assay dependent concentration.
    IHC-P
    1/1000 - 1/5000.
    Notes
    ELISA
    Use at an assay dependent concentration.
    WB
    Use at an assay dependent concentration. Predicted molecular weight: 161 kDa.

    Not recommended for use under denaturing conditions.

    IP
    Use at an assay dependent concentration.
    IHC-P
    1/1000 - 1/5000.

    Target

    • Function

      Type IV collagen is the major structural component of glomerular basement membranes (GBM), forming a 'chicken-wire' meshwork together with laminins, proteoglycans and entactin/nidogen.
      Arresten, comprising the C-terminal NC1 domain, inhibits angiogenesis and tumor formation. The C-terminal half is found to possess the anti-angiogenic activity. Specifically inhibits endothelial cell proliferation, migration and tube formation. Inhibits expression of hypoxia-inducible factor 1alpha and ERK1/2 and p38 MAPK activation. Ligand for alpha1/beta1 integrin.
    • Tissue specificity

      Highly expressed in placenta.
    • Involvement in disease

      Defects in COL4A1 are a cause of brain small vessel disease with hemorrhage (BSVDH) [MIM:607595]. Brain small vessel diseases underlie 20 to 30 percent of ischemic strokes and a larger proportion of intracerebral hemorrhages. Inheritance is autosomal dominant.
      Defects in COL4A1 are the cause of hereditary angiopathy with nephropathy aneurysms and muscle cramps (HANAC) [MIM:611773]. The clinical renal manifestations include hematuria and bilateral large cysts. Histologic analysis revealed complex basement membrane defects in kidney and skin. The systemic angiopathy appears to affect both small vessels and large arteries.
      Defects in COL4A1 are a cause of porencephaly familial (PCEPH) [MIM:175780]. Porencephaly is a term used for any cavitation or cerebrospinal fluid-filled cyst in the brain. Porencephaly type 1 is usually unilateral and results from focal destructive lesions such as fetal vascular occlusion or birth trauma. Type 2, or schizencephalic porencephaly, is usually symmetric and represents a primary defect or arrest in the development of the cerebral ventricles.
    • Sequence similarities

      Belongs to the type IV collagen family.
      Contains 1 collagen IV NC1 (C-terminal non-collagenous) domain.
    • Domain

      Alpha chains of type IV collagen have a non-collagenous domain (NC1) at their C-terminus, frequent interruptions of the G-X-Y repeats in the long central triple-helical domain (which may cause flexibility in the triple helix), and a short N-terminal triple-helical 7S domain.
    • Post-translational
      modifications

      Lysines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in all cases and bind carbohydrates.
      Prolines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.
      Type IV collagens contain numerous cysteine residues which are involved in inter- and intramolecular disulfide bonding. 12 of these, located in the NC1 domain, are conserved in all known type IV collagens.
      The trimeric structure of the NC1 domains is stabilized by covalent bonds between Lys and Met residues.
      Proteolytic processing produces the C-terminal NC1 peptide, arresten.
    • Cellular localization

      Secreted > extracellular space > extracellular matrix > basement membrane.
    • Target information above from: UniProt accession P02462 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 282191 Cow
      • Entrez Gene: 317711 Cow
      • Entrez Gene: 407107 Cow
      • Entrez Gene: 508632 Cow
      • Entrez Gene: 511602 Cow
      • Entrez Gene: 1282 Human
      • Entrez Gene: 1284 Human
      • Entrez Gene: 1285 Human
      • Entrez Gene: 1286 Human
      • Entrez Gene: 1287 Human
      • Omim: 120070 Human
      • Omim: 120090 Human
      • Omim: 120130 Human
      • Omim: 120131 Human
      • Omim: 303630 Human
      • SwissProt: P02462 Human
      • SwissProt: P08572 Human
      • SwissProt: P29400 Human
      • SwissProt: P53420 Human
      • SwissProt: Q01955 Human
      • Unigene: 17441 Human
      see all
    • Alternative names

      • Arresten antibody
      • BSVD antibody
      • CO4A1_HUMAN antibody
      • COL4A1 antibody
      • COL4A1 NC1 domain antibody
      • COL4A2 antibody
      • COL4A3 antibody
      • COL4A4 antibody
      • COL4A5 antibody
      • collagen alpha-1(IV) chain antibody
      • Collagen IV Alpha 1 Polypeptide antibody
      • Collagen IV Alpha 2 Polypeptide antibody
      • Collagen Of Basement Membrane Alpha 1 Chain antibody
      • Collagen Of Basement Membrane Alpha 2 Chain antibody
      • Collagen Type IV Alpha 1 antibody
      • collagen type IV alpha 1 chain antibody
      • Collagen Type IV Alpha 2 antibody
      • Collagen Type IV Alpha 3 antibody
      • Collagen Type IV Alpha 4 antibody
      • Collagen Type IV Alpha 5 antibody
      • RATOR antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Collagen IV antibody (ab6581)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Collagen IV antibody (ab6581)

      Immunohistochemical analysis of formalin-fixed paraffin-embedded human tissue sections, labelling Collagen IV with ab6581 at a concentration of 10 µg/mL for 1 hour at room temperature. The left panel is human kidney sections with the right panel being human liver sections. Antigen retrival was performed with 0.01 M sodium citrate buffer at pH 6.0 at 99°C for 20 mins. The secondary used was a rabbit peroxidase secondary antibody at a 1/10,000 dilution incubated for 45 mins at room temperature. Counterstaining against nuclear DNA was hematoxylin.

    Protocols

    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab6581? Please let us know so that we can cite the reference in this datasheet.

    ab6581 has been referenced in 7 publications.

    • Kargar-Abarghouei E  et al. Characterization, recellularization, and transplantation of rat decellularized testis scaffold with bone marrow-derived mesenchymal stem cells. Stem Cell Res Ther 9:324 (2018). PubMed: 30463594
    • Hassanpour A  et al. Decellularized human ovarian scaffold based on a sodium lauryl ester sulfate (SLES)-treated protocol, as a natural three-dimensional scaffold for construction of bioengineered ovaries. Stem Cell Res Ther 9:252 (2018). PubMed: 30257706
    • Buno KP  et al. In Vitro Multitissue Interface Model Supports Rapid Vasculogenesis and Mechanistic Study of Vascularization across Tissue Compartments. ACS Appl Mater Interfaces 8:21848-60 (2016). PubMed: 27136321
    • Zhou J  et al. Qianliening capsules influence the apoptosis of benign prostatic hyperplasia epithelial-1 cells by regulating the extracellular matrix. Mol Med Rep 11:3734-40 (2015). PubMed: 25592406
    • Whittington CF  et al. Collagen-polymer guidance of vessel network formation and stabilization by endothelial colony forming cells in vitro. Macromol Biosci 13:1135-49 (2013). IHC . PubMed: 23832790
    • Vandenbroucke RE  et al. Matrix metalloprotease 8-dependent extracellular matrix cleavage at the blood-CSF barrier contributes to lethality during systemic inflammatory diseases. J Neurosci 32:9805-16 (2012). IHC ; Mouse . PubMed: 22815495
    • Thomas SN  et al. Impaired humoral immunity and tolerance in K14-VEGFR-3-Ig mice that lack dermal lymphatic drainage. J Immunol 189:2181-90 (2012). PubMed: 22844119

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-7 of 7 Abreviews or Q&A

    ELISA abreview for Anti-Collagen IV antibody (Biotin)

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Purified protein (collagen IV)
    Specification
    collagen IV
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
    Type
    Sandwich (Detection)
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 16 2017

    Question

    Is that under denaturing conditions? 400KDA is a very high molecular weight normally it is hard to get it into tris glycine electrophoresis gels unless we reduce it first with BME or DTT reducing agents. Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 21 2005

    Answer

    Thank you for your enquiry. Yes, 400kDa is very large. May I advise you use lower percentage gels, i.e. 5% or 4-8% gradient. I know it is not easy. We recommend non-denaturing, non-dissociating SDS-PAGE. I have been informed that there is considerable conflicting data in the literature when Collagens are reduced. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Sep 22 2005

    Question

    What is the molecular weight of this protein as detected by Western blot?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 20 2005

    Answer

    Thank you for your enquiry. The native collagen IV is about 400kDa following detection by western blot. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Sep 21 2005

    Question

    Customer would like to know the concentration, if possible, of this ab. Also, she received 10 ul in her vial and is concerned b/c that seems low.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 02 2004

    Answer

    The concentration of the lot that you received is 2.15 mg/ml and you should have received 47 ul in the vial. I apologize for this shortage and will have another vial sent to you free of charge. Can you send me your shipping address including phone number? Also, please tell me what the original Abcam order number was.

    Read More

    Abcam Scientific Support

    Answered on Mar 03 2004

    Question

    AB6581-100 Please let me know the crossreactivity with rat? Best regards, Jonas Hink

    Read More

    Abcam community

    Verified customer

    Asked on Oct 09 2001

    Answer

    This antibody has been tested on various mammalian tissue (although not rat) and shown to work. Given the conservation of the collagen famiily we predict that this antibody will work well on rats.

    Read More

    Abcam Scientific Support

    Answered on Oct 09 2001

    Question

    Please inform me the concentration of immunoglobulin and the volume of liquid.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 03 2001

    Answer

    The protein concnetration is 1mg/ml and the vial size is 0.1ml.

    Read More

    Abcam Scientific Support

    Answered on Aug 03 2001

    Question

    Hello, #AB6581-100(Anti collagen type IV) Could you please let us know the crossreactivity with mouse? I look forward to receiving your early reply. Best regards, Maki Kobayashi

    Read More

    Abcam community

    Verified customer

    Asked on Jul 11 2001

    Answer

    The anti-collagen antibody should cross-react with mouse.

    Read More

    Neil Hayward

    Abcam Scientific Support

    Answered on Jul 12 2001

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