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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Collagen IV antibody [EPR20966] (ab214417)

Submit a review Submit a question References (5)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] (ab214417)
  • Western blot - Anti-Collagen IV antibody [EPR20966] (ab214417)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] (ab214417)
  • Anti-Collagen IV antibody [EPR20966] (ab214417)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20966] to Collagen IV
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Collagen IV antibody [EPR20966]
    See all Collagen IV primary antibodies

  • Description

    Rabbit monoclonal [EPR20966] to Collagen IV

  • Host species

    Rabbit

  • Specificity

    The immunogen is within the Arresten region of Collagen alpha-1(IV) chain.

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human kidney and thyroid carcinoma tissues.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab214417 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
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 200 kDa (predicted molecular weight: 161 kDa).
Notes
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 200 kDa (predicted molecular weight: 161 kDa).

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

    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) - Anti-Collagen IV antibody [EPR20966] (ab214417)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] (ab214417)

    Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue labeling Collagen IV with ab214417 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Basement membrane staining on human thyroid cancer. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Collagen IV antibody [EPR20966] (ab214417)
    Western blot - Anti-Collagen IV antibody [EPR20966] (ab214417)
    All lanes : Anti-Collagen IV antibody [EPR20966] (ab214417) at 1/500 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : Col4a1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 161 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-Collagen IV antibody [EPR20966] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab214417 was shown to bind specifically to Collagen IV. A band was observed at 200 kDa in wild-type HeLa cell lysates with no signal observed at this size in Col4a1 knockout cell line ab255379 (knockout cell lysate ab263755). To generate this image, wild-type and Col4a1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] (ab214417)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen IV antibody [EPR20966] (ab214417)

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Collagen IV with ab214417 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Basement membrane staining on human kidney (PMID: 20551626, PMID: 24445937). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-Collagen IV antibody [EPR20966] (ab214417)
    Anti-Collagen IV antibody [EPR20966] (ab214417)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (5)

Publishing research using ab214417? Please let us know so that we can cite the reference in this datasheet.

ab214417 has been referenced in 5 publications.

  • Li Z  et al. MicroRNA-133b alleviates doxorubicin-induced cardiomyocyte apoptosis and cardiac fibrosis by targeting PTBP1 and TAGLN2. Int J Mol Med 48:N/A (2021). PubMed: 33982775
  • Oliver E  et al. Self-organising human gonads generated by a Matrigel-based gradient system. BMC Biol 19:212 (2021). PubMed: 34556114
  • Sun J  et al. MiR-325-3p inhibits renal inflammation and fibrosis by targeting CCL19 in diabetic nephropathy. Clin Exp Pharmacol Physiol 47:1850-1860 (2020). PubMed: 32603491
  • Shi Q  et al. Interplay between RNA-binding protein HuR and Nox4 as a novel therapeutic target in diabetic kidney disease. Mol Metab 36:100968 (2020). PubMed: 32240965
  • Mou F & Mou C Necrostatin-1 Alleviates Bleomycin-Induced Pulmonary Fibrosis and Extracellular Matrix Expression in Interstitial Pulmonary Fibrosis. Med Sci Monit 26:e919739 (2020). PubMed: 32019905

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