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:
  • Hu YS  et al. Self-assembling vascular endothelial growth factor nanoparticles improve function in spinocerebellar ataxia type 1. Brain 142:312-321 (2019). Read more (PubMed: 30649233) »
  • Deng J  et al. Protective effect of rosiglitazone on chronic renal allograft dysfunction in rats. Transpl Immunol N/A:N/A (2019). Read more (PubMed: 30682409) »
See all 91 Publications for this product

Customer reviews and Q&As

11-20 of 26 Abreviews or Q&A

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.

Read More

Answer

Thanks for your enquiry. ab19808 is diluted in deionized water. I hope this is helpful. Please contact us again if you have any further questions.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Kidney)
Specification
Kidney
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 09 2010

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain blood vessels)
Specification
brain blood vessels

Ms. Christina Whiteus

Verified customer

Submitted Oct 20 2010

Application
Western blot
Sample
Mouse Tissue lysate - whole (pancreatic cancer tissue sample)
Loading amount
25 µg
Specification
pancreatic cancer tissue sample
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 07 2010

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (pancreatic cancer associated collagen deposit in s)
Specification
pancreatic cancer associated collagen deposit in s
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 06 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse ear vessel)
Specification
Mouse ear vessel
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100 0,1%
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C

Abcam user community

Verified customer

Submitted Jun 04 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (skin)
Specification
skin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Dr. Pawel Mazur

Verified customer

Submitted Jun 03 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Bone marrow cells)
Specification
Bone marrow cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 27 2010

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Cell (tooth)
Specification
tooth
Fixative
frozen
Permeabilization
No
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 20°C

Dr. Adnane Nait Lechguer

Verified customer

Submitted Mar 16 2010

11-20 of 26 Abreviews or Q&A

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