概述

  • 产品名称Anti-Collagen IV抗体
    参阅全部 Collagen IV 一抗
  • 描述
    兔多克隆抗体to Collagen IV
  • 特异性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.
  • 经测试应用适用于: ELISA, IHC-Fr, IP, WB, ICC/IF, IHC-P, IHC-FrFl, IHC-FoFrmore details
  • 种属反应性
    与反应: Mouse, Rat, Hamster, Cow, Dog, Human, Pig, Zebrafish, African Green Monkey, Chinese Hamster, Syrian Hamster
    预测可用于: all Mammals
  • 免疫原

    Full length native protein (purified) corresponding to Human Collagen IV aa 1-1669. Collagen Type IV from human and bovine placenta.
    Database link: P02462

  • 阳性对照
    • human epidermal keratinocytes lysate
  • 常规说明

    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.

     

    This antibody is 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.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • 存储溶液pH: 8.00
    Preservative: 0.01% Sodium azide
    Constituents: 4.7625% Sodium borate, 0.146% EDTA, 0.435% Sodium chloride
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 纯化说明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说明This antibody is 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.
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab6586 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ELISA 1/5000 - 1/50000.
IHC-Fr 1/50 - 1/200.
IP 1/100.
WB 1/1000 - 1/10000. Use under non reducing condition. This product is not recommended for use under denaturing conditions in WB, IP, and ELISA. We would suggest testing it under native conditions.
ICC/IF Use at an assay dependent concentration. PubMed: 19933193
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-FrFl Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Highly expressed in placenta.
  • 疾病相关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.
  • 序列相似性Belongs to the type IV collagen family.
    Contains 1 collagen IV NC1 (C-terminal non-collagenous) domain.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Secreted > extracellular space > extracellular matrix > basement membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Arresten 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 2 antibody
    • Collagen Type IV Alpha 3 antibody
    • Collagen Type IV Alpha 4 antibody
    • Collagen Type IV Alpha 5 antibody
    • Collagen Type IV antibody
    see all

Anti-Collagen IV antibody 图像

  • ab6586 staining Collagen IV in Dog liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a 10mM citrate buffer, pH 6.0. Samples were incubated with primary antibody (1/200 in PBS + 1x casein) for 1 hour at 37°C. An undiluted HRP-conjugated Horse anti-rabbit IgG polyclonal was used as the secondary antibody.

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  • Immunocytochemical analysis of Mouse 3T3 cell labeling Collagen IV with ab6586 at 1/250 dilution

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  • Anti-Collagen IV antibody (ab6586) at 1/1000 dilution + Baby Hamster Kidney fibroblasts at 100 µg

    Secondary
    HRP-conjugated donkey anti-rabbit polyclonal IgG at 1/4000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 300 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk

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  • ab6586 staining rat brain tissue sections (ab4616) by IHC-Fr.  Sections were acetone fixed and blocked with 1% serum for 20 minutes at 25°C.  The primary antibody was diluted 1/300 and incubated with the sample for 30 minutes at 25°C.  A biotinylated goat anti-rabbit IgG antibody was used as the secondary.

     

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  • ab6586 staining Collagen IV in heart tissue by Immunohistochemistry (Frozen sections). The sections were fixed in Acetone prior to blocking with 100% SuperBlock Blocking Buffer for 20 mins at 23°C. The primary antibody was diluted 1/50 and incubated with the sample for 12 hours at 4°C. ab6720 was used as the secondary antibody, diluted 1/100.

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  • ab6586 staining Collagen IV in pig epithelial tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in acetone then incubated with ab6586 at a 1/200 dilution for 1 hour at 25°C. The secondary used was ab96886, a goat polyclonal to rabbit IgG - H&L (DyLight® 649), used at a 1/500 dilution.

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  • ab6586 staining Collagen IV in Human corneal epithelial tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was enzymatic using trypsin. Samples were incubated with primary antibody (1/500 inPBS + 2%NGS) for 18 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal (1/500) (ab150077) was used as the secondary antibody.

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  • All lanes : Anti-Collagen IV antibody (ab6586) at 1/1000 dilution

    Lane 1 : Mouse muscle whole tissue lysate
    Lane 2 : Mouse muscle whole tissue lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP conjugated Goat anti-rabbit IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 200 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    This image is courtesy of an anonymous Abreview

    Blocked with 5% BSA for 1 hour at 25°C.

    See Abreview

Anti-Collagen IV antibody (ab6586)参考文献

This product has been referenced in:
  • Löffler T  et al. Decreased Plasma Aß in Hyperlipidemic APPSL Transgenic Mice Is Associated with BBB Dysfunction. Front Neurosci 10:232 (2016). IHC-Fr ; Mouse . Read more (PubMed: 27313503) »
  • Jiang M  et al. Changes in tension regulates proliferation and migration of fibroblasts by remodeling expression of ECM proteins. Exp Ther Med 12:1542-1550 (2016). WB ; Human . Read more (PubMed: 27588075) »

See all 92 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rabbit Cell lysate - whole cell (Rabbit Reticulocyte)
Gel Running Conditions Reduced Denaturing
Loading amount 0.5 µg
Specification Rabbit Reticulocyte
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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提交于 Nov 21 2016

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (A549 (human lung epithelium cells))
Total protein in input 5e+006 cells
Immuno-precipitation step Other - Protein G Dynabeads
Specification A549 (human lung epithelium cells)
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提交于 Nov 04 2016

Application Immunoprecipitation
Sample Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Total protein in input 5e+006 cells
Immuno-precipitation step Other - Protein G Dynabeads
Specification Mouse embryonic fibroblasts (MEFs)
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提交于 Nov 04 2016

Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Brain)
Permeabilization Yes - 0.3 % Triton X-100 included in blocking step
Specification Brain
Blocking step 10 % Donkey serum, 3 % BSA, 0.3 % triton X-100 in PBS as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative Paraformaldehyde
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提交于 Sep 13 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Sheep Tissue sections (artery)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization No
Specification artery
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative 10% normal buffered formalin
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提交于 Sep 01 2016

Application Western blot
Sample Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 50000 cells
Specification BHK-21 (Kidney Fibroblasts)
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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提交于 Jul 22 2016

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (VERO E6)
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 75000 cells
Specification VERO E6
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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提交于 Jul 20 2016

I looked at the alignment between the Collagen IV from Hydra vulgaris (http://www.uniprot.org/uniprot/V9GW26) and the immunogen used to raise ab6586, the alignment is only 42%.
Therefore, I would not recommend this antibody for Hydra vulgaris sampl...

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Application Immunocytochemistry
Sample Human Cultured Cells (Human Breast)
Permeabilization Yes - Triton X-100
Specification Human Breast
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 25°C
Fixative Paraformaldehyde
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提交于 Apr 05 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: MW pH9
Permeabilization No
Specification Brain
Blocking step Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 1500µg/mL · Temperature: RT°C
Fixative Paraformaldehyde
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Gaia Brezzo

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提交于 Jan 29 2016

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