Overview

  • Product name
    Anti-Collagen IV antibody [COL-94]
    See all Collagen IV primary antibodies
  • Description
    Mouse monoclonal [COL-94] to Collagen IV
  • Host species
    Mouse
  • Tested applications
    Suitable for: ELISA, IHC-FoFr, IHC-P, Dot blot, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Non human primatesDoes not react with: Rat, Sheep, Goat, Chicken, Cat
  • Immunogen

    Full length native protein (purified) (Human).

  • Epitope
    Recognises an epitope located on the alpha 1 and/or alpha 2 chains of human collage type IV.
  • Positive control
    • Natural Human Collagen IV protein (ab7536) can be used as a positive control in WB. Human skin tissue.
  • General notes

    Collagen IV is a major constituent of the basement membranes along with laminins and enactins. It is composed of alpha 1 IV chain and alpha 2 IV chain in 2:1 ratio . It can form insoluble fibers with high tensile strength. Antibody to collagen IV is useful in detecting the loss of parts of basement membrane in carcinomas.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab6311 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
ELISA Use at an assay dependent concentration.
IHC-FoFr 1/500.
IHC-P 1/25 - 1/50. Perform enzymatic antigen retrieval before commencing with IHC staining protocol. PubMed: 18843029

We suggest using an ABC detection system.

Dot blot Use at an assay dependent concentration.
ICC/IF 1/200.
IHC-Fr 1/500.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovary tissue sections labeling Collagen IV with ab6311 at 1/400 dilution. Tissue was blocked with 1.4% serum for 30 minutes. Heat mediated antigen retrieval was performed. An undiluted monoclonal alkaline phosphatase conjugated secondary antibody was used.
    Blood vessels in whole ovarian section. Clear staining is seen in the vessel walls.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of human kidney cells labeling Collagen IV with ab6311 at 1/100 dilution. Cells were fixed with paraformaldehyde and blocked with 5% BSA for 30 minutes at 25°C. A polyclonal rabbit anti-mouse Alexa Fluro® 488 was used as the secondary antibody at 1/200 dilution.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labeling Collagen IV with ab6311.

  • Immunofluorescence analysis of Human endothelial cells, staining Collagen IV with ab6311.
    Cells were fixed with formaldehyde and permeabilized with acetone. Samples were incubated with primary antibody (0.5 µg/ml in diluent) for 18 hours at 4°C. An AlexaFluor®488-conjugated goat anti-mouse polyclonal IgG (5 µg/ml) (ab150113) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Application
ELISA
Sample
Human Purified protein (collagen IV)
Specification
collagen IV
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Type
Sandwich (Capture)

Patricia Carulla

Verified customer

Submitted Aug 17 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 08 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.05% typsin
Sample
Human Tissue sections (human whole fat tissue)
Specification
human whole fat tissue
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 18 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (kidney)
Specification
kidney
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 11 2015

Answer

Thank you for contacting us.

Thank you for clarifying that you would prefer to do IHC on paraffin tissue sections rather than frozen tissue sections. For both of these applications I would indeed suggest checking the biocompare website. Ideally you would need a mouse anti-rabbit collagen IV antibody. If this is not possible, then I would try looking for a mouse anti-human collagen IV that has been proven to react with rabbit collagen IV since unfortunately, we do not have this in our catalogue.

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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Answer

Thank you for contacting us.

I am sorry, I was under the impression that you wanted to do immunohistochemistry on rabbit tissue. If this is not the case, what application would you like to use the anti-collagen IV antibody in?

With regards to finding an antibody which is likely to work rabbit, I have managed to find the protein sequences of the alpha chain 1,2 and 4 of rabbit collagen IV at:

http://www.uniprot.org/uniprot/Q7M2X2
http://www.uniprot.org/uniprot/Q7M2X5
http://www.uniprot.org/uniprot/P55787

respectively.

However, some of these sequences are incomplete. For the amino acid data we do have, it appears that there is a relatively high % homology between rabbit and human however because, our data is incomplete and it appears that the rabbit protein may be smaller, I am unable to predict whether an antibody is likely to work or not in rabbit.

I would like to recommend checking the Biocompare website which has an excellent antibody search facility that includes many suppliers to see if they have an anti-collagen IV predicted to work in rabbit. The links are:
www.biocompare.com
http://www.biocompare.com/ProductCategories/2045/Antibodies.html

Otherwise, if you would like to test one of the antibodies from our catalogue we have 7 mouse monoclonals which are guaranteed to work in IHC to choose from. If you would like to test one of them, please contact us letting us know which one you would like to test.

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

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Endothelial in Matrigel)
Specification
Endothelial in Matrigel
Fixative
Formaldehyde
Permeabilization
Yes - Acetone

Abcam user community

Verified customer

Submitted Sep 25 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (skin)
Specification
skin
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.2% Trititon X-100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 17 2012

Answer

Thank you for your reply. I am very pleased to hear you would like to accept our offer and test ab6311 in mouse.

DISCOUNT CODE: 100%ABR-E76ZQ
Expiration date:13/11/11

This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview formouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question

we will be interested by buying AB6311. But I must specify that we want to use the antibody for Immunofluorescence. We are cultivating human cells that are producing collagen IV in a mouse collagen IV matrice. We want to distinguish the human collagen to the mouse one. We expect the human one to be localize around the cells, where the mouse one will recover the whole substrate.

If this will meet the requirement for a test, we will be glad to participate to the collaboration program with you.

Aurelien Forget



Ph-D student

Universitaet Albert-Ludwig

BIOSS

Institut fur Makromolekular Chemie

Stefan-Meier-Str.31

79104 Freiburg

GERMANY

Tel: 00 49 761 203 6310

email: mailto:aurelien.forget@bioss.uni-freiburg.de





On Jul 6, 2012, at 2:30 PM, mailto:technical@abcam.com wrote:















https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header








Dear Aurelien



Thank you for contacting us in regards to anti-Collagen IV antibodies.

As discussed over the phone, we do not currently have any antibodies which have been shown to detect human Collagen IV but not the mouse protein in Western blotting (or any other application). Unfortunately, as all the antibodies which might be applicable have been raised against the full length protein it is not possible to tell how likely it is that it will cross react.

As we discussed, ab6311, has been shown to not cross react with a number of different species but this does not necessarily mean that it will not cross react with mouse. However, as it is a monoclonal antibody, if it does not cross react it is likely to show this specificity consistently from batch to batch and may therefore be most worthwhile trying.

If you would like to try the antibodyab6311 or the other mouse monolconal ab86042 in Western blotting we would be able to offer you a testing discount.

This offer means that if you test the antibodywith mousesamples inWestern blotting and share the results with us through an Abreview (no matter whether positive or negative) you would be eligible for a free primary antibody of your choice from our catalogue (or the equivalent price off any product in our catalogue). More information on this offer can be found here:

https://www.abcam.com/collaboratordiscount

The only constraint to this offer is that the antibody to be tested must be purchased, tested, the Abreview submitted and the free primary antibody claimed within a 4 month period.

When performing the western blot, it would be very helpful if you could show both the positive and negative data (i.e. blotting with the human sample alongside the mouse sample to make sure the antibody is working effectively).

If you would like to participate in this scheme please do let me know as a discount code needs to be issued prior to purchasing the antibody. Please also let me know which of the two antibodies you would like to test (ab6311 or ab86042).

I look forward to receiving your reply.

Help us improve our service.
https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3928259


Best regards,
Karin

Karin Nordgren
Scientific Support Specialist
Abcam plc
https://www.abcam.com


Your original inquiry to Abcam:


Phone call requesting information on antibodies against Collagen IV able to differentiate between mouse and human target.








Abcam Customer Services and Scientific Support Team
https://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body

[CCE3928259]









Discover moreathttp://abcam.com

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Answer

Thank you for your reply and further enquiry which has been forwarded to me as my colleague Karin is currently away from the office.

As Karin previously discussed with you, ab6311, has been shown to not cross react with a number of different species but this does not necessarily mean that it will not cross react with mouse. It is regrettably very difficult to suggest antibodies to customers that we can guarantee to work in one species and definitely not in another. Some antibodies can even show inconsistent results in one particular species, so could work for one customer and not another.

Therefore, I can offer a discount off a future purchase if you buyab6311 now totest in mouse and submit feedback to us in the form of an Abreview. This is on the understanding that we cannot guarantee this antibody willnot detect in mouse.

For us, it does not matter whether the Abreview is positive or negative, we would just really like to receive your feedback of the results in mouse. The discount would be to the value of: 1 free primary antibody.

If you are interested in this offer, please I would appreciate if you could take time to read through the terms and conditions and then follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab6311 in mouse. I will then send a discount code. This code must be issued before purchasing ab6311 so please wait for my reply before ordering.

2. Purchase ab6311 either by phone, fax, or online (www.abcam.com).

3. Test it in mouse.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated!Whether ab6311 turns out to be suitable orunsuitable for detection of Collagen IV in mouse, you will still receive the discount on your next purchase after your Abreview has been submitted.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Please let me know if you have any questions and I would be happy to help you further. I look forward to hearing from you with confirmation that you would like to proceed with the offer.

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1-10 of 22 Abreviews or Q&A

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