Native Human Collagen IV protein (ab7536)

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Overview

Description

  • Nature
    Native
  • Source
    Native
  • Amino Acid Sequence
    • Accession
      P02462
    • Species
      Human
    • Molecular weight
      161 kDa
    • Additional sequence information
      Prepared from human placenta and is chromatographically and immunologically pure.

Specifications

Our Abpromise guarantee covers the use of ab7536 in the following tested applications.

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

  • Applications

    Western blot

    Immunoprecipitation

    ELISA

    Blocking

  • Form
    Liquid
  • Additional notes

    This product is free from other collagens, human serum proteins and non-collagen extracellular matrix proteins. This product reacts with anti-Collagen Type IV. Reaction with anti-Collagen I, II, III, V or VI is negligible (typically less than 1% cross reactivity was detected by ELISA).

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

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

    pH: 4.50
    Preservative: 0.01% Sodium azide
    Constituent: 0.82% Sodium acetate

General Info

  • Alternative names
    • Arresten
    • BSVD
    • CO4A1_HUMAN
    • COL4A1
    • collagen alpha-1(IV) chain
    • collagen type IV alpha 1 chain
    • RATOR
    see all
  • 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

References

This product has been referenced in:
  • Xiong Y  et al. Precursor N-cadherin mediates glial cell line-derived neurotrophic factor-promoted human malignant glioma. Oncotarget 8:24902-24914 (2017). Read more (PubMed: 28212546) »
  • Sand JM  et al. MMP mediated degradation of type IV collagen alpha 1 and alpha 3 chains reflects basement membrane remodeling in experimental and clinical fibrosis--validation of two novel biomarker assays. PLoS One 8:e84934 (2013). ELISA . Read more (PubMed: 24376856) »
See all 4 Publications for this product

Publishing research using ab7536? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Question

Hi,

I would like to coat my samples with collagen IV protein (ab7536). The plates are 24 well plates. Would you please let me know the recommended concentration of collagen IV protein (e.g. ug/cm2 or ug/ml) for coating the samples.

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Answer

Thank you for contacting us. We would recommend using a concentration of 20 ug/ml for coating the plate.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

We have tried to probe collagen IV in samples run in non-denaturing conditions using ab6586, and using ab7536 as a positive control. We have not been very successful, and we would like to know whether you could provide any image showing the results from western blotting in non denaturing conditions using this antibody, so we could know what to expect. Since you recommend to use it in non denaturing conditions I assume that you have some positive results that we use as a reference.

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Answer

Thanks for your reply.

I have searched images in published papers using ab7536 and western blotting data fromuser's online reviews, all associated with the online datasheet for this antibody. Interestingly, I have not beenfound a blot detecting native protein. All blots I have found resolve samples vial denaturing/reducing conditions. I am not familiar with the collagen literature but it is surprising that even published papers use denatured samples. There may be information in the literature illustrating that certain collagens in certain samples can be detected specifically under denaturing conditions.

It is typically more difficult to interpret western blotting data from native samples. Molecular weight is not easily determined as proteins migrate based on size, charge, modification state, etc. And of course, defferent sample preparation methods and different samples may produce different results.

I'd be happy to look at your western blot data if you are interested in forwarding it along as an attachement.

Thanks very much!

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Question


I have two technical questions about the antibody with catalog number ab6586 (rabbit anti-collagen IV).



I have read the posts in the scientific support tab for this antibody, and it is not clear to me whether it is better to run the samples for WB in denaturing conditions or in non-denaturing conditions. I have tried denaturing conditions and the results are confusing, so I wonder if trying the non-denaturing PAGE will provide a different and more reliable result.



My second question is related to your product with catalog number ab7536 (collagen IV protein). We are considering to use this protein as a positive control in our western blots (in non denaturing conditions). Is there any report of previous use of this protein as a positive control in WB? Does the antibody (ab6586) recognize the same epitope in this protein? Also, is there any collagen IV blocking peptide in you catalog that we could use to identify our band?

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Answer

Thanks for your enquiry.

I would definitely recommend running a non-denaturing gel. It is typically recommended to blot for collagen under non-denaturing conditions as the primary structure of collagens are quite similar. Under non denaturing conditions the collagen's 3 dimensional structure is retained and it is this 3 dimensional structure that is recognized by anti-collagen antibodies.

Regarding your second question, ab7536 has been used under non denaturing conditions in western blotting. Unfortunately we do not have any western blotting data available using this protein. There is not specific report of ab6586 being used to detect ab7536 via western blotting but in theory it should work using non denaturing WB conditions.

I hope this is helpful. Please contact us again if you have any further questions.

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Question

What is the molecular weight of this protein? Is it the full protein, containing both the alpha 1 and 2 chains?

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Answer

Thank you for your enquiry. This product is the full protein consisting of both chains. I am having difficulty finding the molecular weight of collagen IV. I found one source that reports the molecular weight as approx. 300kDa. It is difficult to find the MW of the complete protein, because when run on a gel, typically there are many fragments, but the MW of the total protein is approx. 300 kDa. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

How is the concentration ab7536 determined?

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Answer

Thank you for your enquiry. The concentration of ab7536 Collagen Type IV is determined by dry weight.

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Question

My order number is 48702 and I receipt the package Aug, 3. However, when I aliquoted I found the volum of one vial ( product code is ab7536---collagen IV from human placenta) is even less than 300ul, it should be 500ul. The other one is ok. I'll appreciate it if you can send me another vial. Thanks again and have a good day!

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Answer

Thank you for your email. I apologize for the shortage that you received and I am sending you another vial free of charge. For your record it is order# 50349 and you will receive the vial Wednesday. Please let me know if you have any further questions.

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Question

How many bands can be seen on SDS-PAGE eletrophoresis of ab7536? What are their molecular weight? Thanks a lot.

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Answer

The answer to this question depends on several variables, such as source, method of preparation, how they were stored and handled. I have a reference showing three major bands (125kDa, 145kDa and 165kDa) under reduced condition of SDS-PAGE.

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