Overview

  • Product name
    Anti-Collagen IV antibody (Biotin)
    See all Collagen IV primary antibodies
  • Description
    Rabbit polyclonal to Collagen IV (Biotin)
  • 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, IP, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Cow, Human
    Predicted to work with: Mammal
  • Immunogen

    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.

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
    pH: 8.00
    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

Applications

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 1/4000 - 1/8000.
IP Use at an assay dependent concentration.
WB 1/5000 - 1/10000. Not recommended for use under denaturing conditions.
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.
  • Information by UniProt
  • Database links
  • Alternative names
    • Arresten antibody
    • BSVD antibody
    • CO4A1_HUMAN antibody
    • COL4A1 antibody
    • collagen alpha-1(IV) chain antibody
    • collagen type IV alpha 1 chain antibody
    • RATOR antibody
    see all

Images

  • 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.

References

This product has been referenced in:
  • 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). Read more (PubMed: 27136321) »
  • 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 . Read more (PubMed: 23832790) »

See all 4 Publications for this product

Customer reviews and Q&As

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)
Username

Abcam user community

Verified customer

Submitted Aug 16 2017

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 considera...

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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.

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 numb...

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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.

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

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

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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