Overview

  • Product name
  • Description
    Rabbit polyclonal to Collagen IV
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, IHC-FoFr, WB, IHC-FrFl, ICC/IF, RIA, ELISA, IHC - Wholemountmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Full length native protein (extracted and purified from tumor tissues) (Mouse).

  • Positive control
    • IHC-P: Mouse brain, pancreas and mammary gland tissue. ICC/IF: Mouse ovaries, and brain cells. IHC-fr: Mouth tooth tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab19808 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-Fr 1/400.

Permeabilisation with ~0.2% Triton is recommended

IHC-P 1/500.
IHC-FoFr Use at an assay dependent concentration. PubMed: 19762493
WB Use at an assay dependent concentration. PubMed: 23249995
IHC-FrFl Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 22253831
RIA Use at an assay dependent concentration.
ELISA 1/200.
IHC - Wholemount Use at an assay dependent concentration. PubMed: 24353059

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 paraffin-embedded mouse pancreas tissue stained for Collagen IV using ab19808.

    Endocrine-cells coated with a layer of extracellular matrix (P1). Immunohistochemical staining for Insulin (green), glucagon (red) and collagen IV (yellow) is shown. Note that intra-islet blood vessels are also associated with the extracellular matrix. Scale bar is 50 µm.

    The primary antibodies were detected using different combinations of Cy2, Cy5 and Texas Red-conjugated secondary antibodies and the image was captured via confocal microscopy.

  • Ovaries were dissected from PND 23–29 mice, embedded in Cryomatrix and fixed with 4% formaldehyde for 10 minutes, rinsed three times with phosphate-buffered saline (PBS), then permeabilized with 0.1% Triton X-100 for 15 minutes. Sections were again rinsed three times with PBS, blocked for 30 minutes with blocking solution (5% BSA in 0.1% Triton X-100), then rinsed three times with blocking solution. The tissue was then incubated for one hour with ab19808 at 1/400 dilution. Sections were then rinsed three times in blocking solution and incubated in secondary antibody (FITC-conjugated goat anti-rabbit secondary antibody).

    Immunofluorescence with ab19808 was used to detect Collagen IV localization to the follicular basal lamina (white filled arrowhead), focimatrix (open arrowhead), thecal matrix (asterix), and endothelial basal lamina of stromal blood vessels (square).

  • ab19808 staining Collagen IV in murine brain tissue/ human xenograft tissue by Immunohistochemistry. Tissue was fixed in AFA (alcohol-formal-acetate) and a heat mediated antigen retrieval step was performed using Tris-EDTA pH 9. Samples were then blocked using 3% BSA for 30 minutes at 20°C and then incubated with ab19808 at a 1/500 dilution for 1 hour and 30 minutes. The secondary used was an undiluted HRP-conjugated goat polyclonal. Left side: normal mouse brainRight side: human glioblastoma xenograft

    See Abreview

  • ab19808 at a 1/200 dilution staining mouse mammary gland tissue by Immunohistochemistry (Formalin-fixed paraffin-embedded sections). The antibody was incubated with the tissue for 16 hours and then detected with an Alexa Fluor® 488 conjuaged anti-rabbit antibody.

    This image is courtesy of an Abreview submitted by an anonymous researcher on 26 January 2006.

    See Abreview

  • ab19808 staining Collagen IV in mouse brain cells (ab30149) by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with formaldehyde and blocked with 0.25% TNB for 30 minutes at 22°C. Samples were incubated with primary antibody 1/250 in TNB for 18 hours at 22°C. A Biotin-conjugated Goat polyclonal to rabbit IgG (ab6720), dilution 1/500, was used as secondary antibody.

    See Abreview

  • ab19808 staining Collagen IV in mouse tooth tissue section by Immunohistochemistry (Frozen sections). Tissue samples were blocked with 1% BSA for 20 minutes at 200C and incubated with undiluted primary antibody for 1 hour at 200C. An Alexa Fluor®594-conjugated chicken polyclonal to rabbit IgG was used as secondary antibody at 1/500 dilution. Red colour in the image represents staining of CoIlagen-IV and the green is for CD31, yellow for ED16+6, which were purchased from different sources.

    See Abreview

References

This product has been referenced in:
  • Peng J  et al. YAP and TAZ mediate steroid-induced alterations in the trabecular meshwork cytoskeleton in human trabecular meshwork cells. Int J Mol Med 41:164-172 (2018). Read more (PubMed: 29115373) »
  • Luo X  et al. Choroidal pericytes promote subretinal fibrosis after experimental photocoagulation. Dis Model Mech 11:N/A (2018). Read more (PubMed: 29622551) »
See all 82 Publications for this product

Customer reviews and Q&As

1-10 of 25 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (femoral ertery)
Antigen retrieval step
None
Permeabilization
Yes - 0.1% Triton-X
Specification
femoral ertery
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 16 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
Yes - 0.5% Triton
Fixative
Paraformaldehyde

Ms. Diana Cifuentes

Verified customer

Submitted Dec 01 2014

Question
Answer

Unfortunately the concentration is not determined for this product.

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Question
Answer

Thank you for your recent telephone enquiry. i am sorry it has taken some time to obtain this information for you from the originator.

I am sorry to confirm that in this particular case, the buffer components are proprietary. We are able to inform you that it includes a phosphate based salt (not PBS or TBS) which promotes the tissue activity and the quality of images in immunohistochemistry assays. No additional proteins or amines are added to the buffer.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for contacting us.

There isn’t a specific protocol for performing WB with this antibody. We recommend using reducing and denaturing conditions unless otherwise stated on the datasheet. For this particular antibody there isn’t a specific recommended dilution, so I would suggest trying several dilutions to determine the one that works better for your experiment. For purified antibodies, the starting recommended concentration is 1ug/ml.

In case it could be useful, I include the link to our protocol’s page, where you can find all the information relative to Western Blot protocols:

https://www.abcam.com/index.html?pageconfig=popular_protocols

In any case, all of our products are covered by our Abpromise® guarantee, which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

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

Read More

Answer

Thanks for your call today and for your questions about our collagen IV antibodies.
As we discussed, I would still recommend trying ab6586 with your standard Western blotting conditions. We recommend non-denatured conditions because the affinity of the antibody is higher in these conditions, but I did find user-submitted Abreviews in which the antibody was used in denatured conditions and the results were satisfactory (rated "Excellent" by the reviewers)-
https://www.abcam.com/index.html?datasheet=6586&tab=abreviews&intabreviewid=7703
https://www.abcam.com/index.html?datasheet=6586&tab=abreviews&intabreviewid=8531
Please note that the expected band size in these conditions is between 200 and 250 kDa.
I hope this information will be useful, but if you have any further questions or need anything else, please let me know and I'll be happy to help. I apologize for any confusion regarding the information on our datasheet, and I will see if we can update the Western blot application information.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Specification
brain
Fixative
Alcool-Formol-Acetate (AFA)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse brain cells)
Specification
Mouse brain cells
Fixative
Alcool-Formol-Acetate (AFA)
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 15 2011

Answer

Merci de votre réponse. La recette pour le TBS 10x est la suivante : 24.23 g Trizma HCl 80.06 g NaCl Mix in 800 ml ultra pure water. pH to 7.6 with pure HCl. Top up to 1 L. Ceci doit être dilué 10x pour obtenir du TBS 1x. Veuillez trouver les composition de différents tampons aussi sous le lien suivant: https://www.abcam.com/ps/pdf/protocols/Buffers%20and%20stock%20solutions.pdf J'espère que ce protocole va permettre d'observer un beau signal. Je me réjouis de recevoir de vos nouvelles. En attendant, n'hésitez pas à nous recontacter si vous avez des autres questions.

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Answer

Merci pour nous avoir contacté. Je suis désolée que vous avez des problèmes avec un de nos anticorps. Comme discuté, j'ai donc contacté le laboratoire pour avoir plus d'information sur le protocole et le démasquage utilisé avec cet anticorps. En effet, le laboratoire nous à confirmé que les techniques de démasquage à la chaleur sont en générale trop dure pour des études de collagène. Effectivement, celles-ci peuvent induire une altération de la membrane ainsi que une disruption ou encore un bruit de fond élevé. Je vous ai donc mis en document attaché le protocole du laboratoire, qui lui utilise quand même un démasquage mais avec de la glycine et de hyaluronidasse. J'espère que ceci est utile pour vous. Est-ce que vous avez la possibilité d'essayer un traitement enzymatique des lames? Je pense que ceci pourrait être une solution. Je me réjouis d'avoir de vos nouvelles. N'hésitez à me contacter aussi si vous avez des questions.

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1-10 of 25 Abreviews or Q&A

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