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    fibronectin-antibody-ab299.pdf

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Cardiovascular Angiogenesis Adhesion / ECM Extracellular Matrix
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Anti-Fibronectin antibody (ab299)

  • Datasheet
  • SDS
Reviews (2)Q&A (11)References (18)

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Sandwich ELISA - Anti-Fibronectin antibody (ab299)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody (ab299)

Key features and details

  • Rabbit polyclonal to Fibronectin
  • Suitable for: ICC/IF, Sandwich ELISA
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Fibronectin antibody [F14] (ab45688)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Fibronectin antibody
    See all Fibronectin primary antibodies
  • Description

    Rabbit polyclonal to Fibronectin
  • Host species

    Rabbit
  • Specificity

    This antibody reacts with human Fibronectin and has negligible cross-reactivity with Type I, II, III, IV, V or VI Collagens or Laminin. Minimum Cross Reactivity to collagens and non-collagen extracellular matrix proteins.
  • Tested applications

    Suitable for: ICC/IF, Sandwich ELISAmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Human Fibronectin. Fibronectin was purified from Human plasma by binding to a denatured gelatin column followed by elution with high concentrations of arginine. The eluted material was further purified by gel filtration

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.87% Sodium chloride, 0.42% Potassium phosphate
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Prepared by immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against human serum proteins and collagen and non-collagen extracellular matrix proteins to remove any unwanted specificities. This product has been sterile filtered.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Extracellular Matrix
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • ECM Proteins
    • Fibronectin
    • Stem Cells
    • Lineage Markers
    • Ectoderm
    • Stem Cells
    • Neural Stem Cells
    • Extracellular
    • Stem Cells
    • Mesenchymal Stem Cells
    • Osteogenesis
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Extracellular matrix
    • Other
    • Developmental Biology
    • Lineage specification
    • Ectoderm

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Fibronectin type III EDB protein (ab209886)
  • Related Products

    • Native Human Fibronectin protein (ab81743)
    • Native Mouse Fibronectin protein (ab92784)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab299 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
ICC/IF
Use a concentration of 1 µg/ml.
Sandwich ELISA
Use a concentration of 0.5 µg/ml. For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923) as Capture.
Notes
ICC/IF
Use a concentration of 1 µg/ml.
Sandwich ELISA
Use a concentration of 0.5 µg/ml. For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923) as Capture.

Target

  • Function

    Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization. Participates in the regulation of type I collagen deposition by osteoblasts.
    Anastellin binds fibronectin and induces fibril formation. This fibronectin polymer, named superfibronectin, exhibits enhanced adhesive properties. Both anastellin and superfibronectin inhibit tumor growth, angiogenesis and metastasis. Anastellin activates p38 MAPK and inhibits lysophospholipid signaling.
  • Tissue specificity

    Plasma FN (soluble dimeric form) is secreted by hepatocytes. Cellular FN (dimeric or cross-linked multimeric forms), made by fibroblasts, epithelial and other cell types, is deposited as fibrils in the extracellular matrix. Ugl-Y1, Ugl-Y2 and Ugl-Y3 are found in urine.
  • Involvement in disease

    Glomerulopathy with fibronectin deposits 2
  • Sequence similarities

    Contains 12 fibronectin type-I domains.
    Contains 2 fibronectin type-II domains.
    Contains 16 fibronectin type-III domains.
  • Developmental stage

    Ugl-Y1, Ugl-Y2 and Ugl-Y3 are present in the urine from 0 to 17 years of age.
  • Post-translational
    modifications

    Sulfated.
    It is not known whether both or only one of Thr-2064 and Thr-2065 are/is glycosylated.
    Forms covalent cross-links mediated by a transglutaminase, such as F13A or TGM2, between a glutamine and the epsilon-amino group of a lysine residue, forming homopolymers and heteropolymers (e.g. fibrinogen-fibronectin, collagen-fibronectin heteropolymers).
    Phosphorylated by FAM20C in the extracellular medium.
    Proteolytic processing produces the C-terminal NC1 peptide, anastellin.
  • Cellular localization

    Secreted, extracellular space, extracellular matrix.
  • Target information above from: UniProt accession P02751 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2335 Human
    • Omim: 135600 Human
    • SwissProt: P02751 Human
    • Unigene: 203717 Human
    • Alternative names

      • CIG antibody
      • Cold insoluble globulin antibody
      • Cold-insoluble globulin antibody
      • DKFZp686F10164 antibody
      • DKFZp686H0342 antibody
      • DKFZp686I1370 antibody
      • DKFZp686O13149 antibody
      • ED B antibody
      • Fibronectin 1 antibody
      • FINC antibody
      • FINC_HUMAN antibody
      • FN antibody
      • FN1 antibody
      • FNZ antibody
      • GFND antibody
      • GFND2 antibody
      • LETS antibody
      • Migration stimulating factor antibody
      • MSF antibody
      • Ugl-Y3 antibody
      see all

    Images

    • Sandwich ELISA - Anti-Fibronectin antibody (ab299)
      Sandwich ELISA - Anti-Fibronectin antibody (ab299)
      Standard Curve for Fibronectin (Analyte: Fibronectin protein (Human) (ab81743)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923) at 1µg/ml and Detector Antibody Rabbit polyclonal to Fibronectin (ab299) at 0.5µg/ml.
    • Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody (ab299)
      Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody (ab299)
      ICC/IF image of ab299 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab299, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    Protocols

    • Immunocytochemistry & immunofluorescence protocols
    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (18)

    Publishing research using ab299? Please let us know so that we can cite the reference in this datasheet.

    ab299 has been referenced in 18 publications.

    • Yang M  et al. Resveratrol induces PD-L1 expression through snail-driven activation of Wnt pathway in lung cancer cells. J Cancer Res Clin Oncol 147:1101-1113 (2021). PubMed: 33471184
    • Li X  et al. Upregulation of lactate-inducible snail protein suppresses oncogene-mediated senescence through p16INK4ainactivation. J Exp Clin Cancer Res 37:39 (2018). PubMed: 29482580
    • Tanos T  et al. Isolation of putative stem cells present in human adult olfactory mucosa. PLoS One 12:e0181151 (2017). PubMed: 28719644
    • Li S  et al. Hydrogels with precisely controlled integrin activation dictate vascular patterning and permeability. Nat Mater 16:953-961 (2017). PubMed: 28783156
    • Malik M  et al. Gonadotropin-releasing hormone analogues inhibit leiomyoma extracellular matrix despite presence of gonadal hormones. Fertil Steril 105:214-24 (2016). PubMed: 26409322
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 13 Abreviews or Q&A

    Western blot abreview for Anti-Fibronectin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Melanoma cell line)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    15 µg
    Specification
    Melanoma cell line
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 26 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Fibronectin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Sheep Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    No
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 15 2017

    Question

    Please recommend a pair of antibodies and the antigen that will give a strong signal in sandwich ELISA. The antigen can be anything, we just want a strong signal. We may want to biotinylate the capture antibody, and we may need a gold-conjugated secondary antibody for detection.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 22 2013

    Answer

    I found a good set of products that gives a strong sandwich ELISA signal.

    Capture: ab80923

    Detection: ab299

    Antigen: ab81743

    Click here (or use the following: https://www.abcam.com/Fibronectin-antibody-HFN-71-ab80923.html).

    Click here (or use the following: https://www.abcam.com/Fibronectin-antibody-ab299.html).

    Click here (or use the following: https://www.abcam.com/Native-Human-Fibronectin-protein-ab81743.html).

    The capture antibody can be easily biotinylated with one of our EasyLink kits. They come in three sizes:

    Click here (or use the following: https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-3-x-100g-Biotin-ab102864.html).

    Click here (or use the following: https://www.abcam.com/Biotin-type-A-Conjugation-Kit-ab102865.html).

    Click here (or use the following: https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-1-x-1mg-Biotin-ab102863.html).

    If you need an anti-rabbit IgG secondary antibody conjugated to gold, whe have several of these in three particle sizes: 6, 12, and 18 nm. For an example, please see the datasheet for ab105296:

    Click here (or use the following: https://www.abcam.com/Donkey-Rabbit-IgG-HL-18nm-Gold-preadsorbed-ab105296.html).

    Read More

    Abcam Scientific Support

    Answered on Feb 22 2013

    Question

    thank you very much for your email and for the suggestions. However, for our experiments we are looking for a protein that is released into the extracellular matrix of the body. If I am not mistaken OCT-4 seems to stay within the cell. Could you recommend a combination of capture-, detection- and fluorescently labeled second antibody for a protein which is released into the extracellular matrix?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 20 2012

    Answer


    I've included a list of capture, detection, protein, and fluorescent-conjugated secondary below against Fibronectin. The capture and detection antibodies have been tested and guaranteed to work in sandwich ELISA to detect human fibronectin:

    Capture: ab80923 https://www.abcam.com/fibronectin-antibody-hfn-7-1-bsa-and-azide-free-ab80923.html
    Detection: ab299 https://www.abcam.com/fibronectin-antibody-ab299.html
    Fibronectin protein: ab81743 https://www.abcam.com/fibronectin-protein-human-ab81743.html
    Fluorescent-conjugated secondary: any anti-rabbit fluorescent-conjugated IgG secondary, list https://www.abcam.com/Search?AppliedFacets.FacetProductType=Secondary+antibodies&AppliedFacets.FacetTargetName=IgG&AppliedFacets.FacetConjugate=DyLight%C2%AE+488&AppliedFacets.FacetConjugate=DyLight%C2%AE+550&AppliedFacets.FacetConjugate=DyLight%C2%AE+594&AppliedFacets.FacetConjugate=Chromeo%E2%84%A2+546&AppliedFacets.FacetConjugate=Cy2+%C2%AE&AppliedFacets.FacetConjugate=Cy3.5+%C2%AE&AppliedFacets.FacetConjugate=Cy5+%C2%AE&AppliedFacets.FacetConjugate=Cy5.5+%C2%AE&AppliedFacets.FacetConjugate=DyLight%C2%AE+650&AppliedFacets.FacetConjugate=FITC&AppliedFacets.FacetConjugate=Phycoerythrin&AppliedFacets.FacetConjugate=SureLight%C2%AE+Allophycocyanin&AppliedFacets.FacetConjugate=Texas+Red+%C2%AE&AppliedFacets.FacetConjugate=TRITC&AppliedFacets.FacetReactivity=Rabbit&IsFacetsOnly=False&PageNumber=1&Keywords%5b0%5d=anti-rabbit&X-Requested-With=XMLHttpRequest

    The specific sandwich ELISA protocol used to validate these 2 primary antibodies using ab81743 Fibronectin protein as the target is attached to this email.

    Read More

    Abcam Scientific Support

    Answered on Dec 20 2012

    Question

    testing ab299 in IHC-P

    Read More

    Abcam community

    Verified customer

    Asked on Apr 11 2012

    Answer

    Thank you for your phone call.


    DISCOUNT CODE: xxx
    Expiration date: xxx

    I am very pleased to hear you would like to accept our offer and test ab299 in IHC-P. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. 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: https://www.abcam.com/collaborationdiscount.

    Read More

    Abcam Scientific Support

    Answered on Apr 11 2012

    Question

    I am looking for an antibody that will stain macrophages/monocytes in formalin fixed- parafin embedded pancreas sections from mouse (Balb/c nude). I would be very grateful indeed if you could recommend a suitable antibody. Please note that a pan-leukocyte marker would be an acceptable alternative to a specific monocyte/macrophage antibody.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 11 2006

    Answer

    Thank you for your enquiry. We have several antibodies suitable for the labelling of macrophages/monocytes. However, unfortunately many of them have not been tested for theur use in mice. We do have an antibody that recognises the mouse F4/80 antigen, a 160kD glycoprotein expressed by murine macrophages; ab6640 F4/80 antibody [CI:A3-1]. This antibody is a rat monoclonal suitable for application in the following applications; FACS, IHC-Fr, IHC-P, IHC-R, IP, RIA, WB. It has received some very favourable reviews from our customers. Given that it is rat as opposed to mouse; this should prevent excessive background from IHC. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Oct 11 2006

    Question

    One of our customers is testing ab292(collagen I) for WB using human sample, but she has problem to test it. (non specific bands with 1:1000) So the customer would like to get some information to optimize the test result. If you have any information (protocol, specially picture),Please let me know. (we've already provided the review and protocol updated your website) in addition, she will test ab299(fibronectin) as well. Please give us protocol and blot picture of ab299.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 16 2004

    Answer

    Thank you for your email. All the information that we have regarding these two antibodies is located on our online datasheets; we do not have any images available at this time. For ab292 I would recommend decreasing the concentration of the primary antibody - for Western blotting we recommend using ab292 at 1:5,000-1:10,000. Also, please note that greatly diminished reactivity and selectivity of these antibodies will result if denaturing and reducing conditions of SDS-PAGE and immunoblotting are used. For ab299, the antibody (immunoblot) shows a 450kDa band non-reduced and 225 kDa band when fully reduced, and a working dilution of 1:4,000 to 1:8,000 of the stock concentration is suggested. Also, please take a look at the reviews located on the datasheet. If you have any more questions, please contact us again.

    Read More

    Abcam Scientific Support

    Answered on Aug 19 2004

    Question

    We are using your Fibronectin antibody (ab299). And we would like to know if that (ab299) or some other of your Fibronectin antibody would be suitable for coating ELISA microtitre plates. And it would be great if you´d have some experiences about the right concentrations. Thank you and hope to hear from you soon.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 24 2003

    Answer

    This antibody has been used in ELISA by using 100uL unlabeled antibody per well at a concentration of 10ug/mL.

    Read More

    Abcam Scientific Support

    Answered on Jul 30 2003

    Question

    1. Has this antibody been used for elisa and if not can it be used for elisa of cellular (ECM) or plasma fibronectin (SECRETED) from rat source. 2. For elisa of cellular ECM proteins, what is the best cell solubilizing agent in general

    Read More

    Abcam community

    Verified customer

    Asked on Jul 17 2002

    Answer

    This antibody has not been tested in the application or species you describe above, therefore we can't guarantee results. Thesit, triton X100 or Tween 20 are classical solubilizing agents that should work well.

    Read More

    Abcam Scientific Support

    Answered on Jul 18 2002

    Question

    Hello: Does this reagent work for immunofluorescence? I need an anti-fibronectin polyclonal antibody urgently.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 21 2002

    Answer

    This antibody, used in the applications listed and with the appropriate fluorescent-label-conjugated-secondary antibody, should perform well in immunofluorescence.

    Read More

    Abcam Scientific Support

    Answered on Jun 24 2002

    1-10 of 13 Abreviews or Q&A

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