Overview

  • Product name

  • Description

    Rabbit polyclonal to Fibronectin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-FoFr, IHC-P, IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Hamster, Cow, Dog, Human, African green monkey, Chinese hamster, Syrian hamster
  • Immunogen

    Fibronectin isolated from a pool of normal human plasma.

  • Positive control

    • IHC-Fr: Mouse colon and embryonic heart tissue. IHC-P: Human kidney tissue. WB: Human colon lysate; HepG2 and NIH/3T3 cell lysate. ICC/IF: HeLa cells.
  • General notes

      

Properties

Applications

Our Abpromise guarantee covers the use of ab2413 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 at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 285 kDa (predicted molecular weight: 262 kDa).
IP Use at an assay dependent concentration.

Used at 1ug/ml for 1 hr 30 min on renal carcinoma cell line 786-0 (see Abreview).

IHC-FoFr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

This antibody may be diluted to a titer of 1/50 - 1/250 in an ABC method. We suggest an incubation period of 30 minutes at room temperature.

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.
  • Information by UniProt
  • Database links

  • 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

  • Paraformaldehyde-fixed frozen section of mouse colon tissue stained for Fibronectin (green) using ab2413 at 1/200 dilution in immunohistochemical analysis, followed by Alexa Fluor® 488 conjugated Goat anti-Rabbit IgG (H+L).

    See Abreview

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue, staining Fibronectin with ab2413.
  • All lanes : Anti-Fibronectin antibody (ab2413) at 1 µg/ml

    Lane 1 : Human colon tissue lysate - total protein (ab30051)
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 262 kDa
    Observed band size: 285 kDa
    why is the actual band size different from the predicted?

  • Paraformaldehyde-fixed frozen section of mouse embryonic heart tissue stained for Fibronectin (green) using ab2413 at 1/100 dilution in immunohistochemical analysis, followed by Alexa Fluor® 488 conjugated Donkey anti-Rabbit IgG.

    Nuclear counterstain: DAPI (blue).

    See Abreview

  • ICC/IF image of ab2413 stained HeLa cells. The cells were 4% PFA 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 (ab2413, 1 µg/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). This antibody also gave a positive IF result in Hek293, HepG2 and MCF7 cells.
  • ab2413 staining Fibronectin in human kidney tissue section by IHC-P (Formalin/PFA-fixed paraffin embedded tissue sections). Tissue sections were incubated with ab2413 at a dilution of 1:250 for one hour. Heat mediated antigen retrieval technique was used with citrate buffer at pH 6.0. DAB staining was done with a biotinylated secondary for 45 min at RT at a concentration of 1:1000.

References

This product has been referenced in:

  • Qin T  et al. Prediction of the mechanisms of action of Shenkang in chronic kidney disease: A network pharmacology study and experimental validation. J Ethnopharmacol 246:112128 (2020). Read more (PubMed: 31386888) »
  • Soung YH  et al. Arrestin Domain Containing 3 Reverses Epithelial to Mesenchymal Transition and Chemo-Resistance of TNBC Cells by Up-Regulating Expression of miR-200b. Cells 8:N/A (2019). Read more (PubMed: 31295851) »
See all 283 Publications for this product

Customer reviews and Q&As

1-10 of 10 Q&A

Answer

The following antibodies we can recommend as per your requirements. These are tested in IHC-P and WB with mouse samples and are guaranteed.
collagen IV

https://www.abcam.com/collagen-iv-antibody-ab6586.html

anti Laminin

https://www.abcam.com/laminin-antibody-ab128053.html

anti Fibronectin
https://www.abcam.com/fibronectin-antibody-ab2413.html

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Answer

When transferring large proteins precipitation or incomplete transfer of the high MW protein can occur. I have included some tips about large protein transfer from our Western Blot guide below and have links this as well. I hope this is of help to you. Note on transfer of large and small proteins The balance of SDS and methanol in the transfer buffer, protein size, and gel percentage can affect transfer efficiency. The following modifications will encourage efficient transfer. Large proteins (>100 kD) For large proteins, transfer out of the gel may be very slow, just as they run slowly within the gel during separation. If blotting a large protein, be sure to run your samples in a low-concentration gel, 8% or less. These will be very fragile, so handle carefully. Large proteins will tend to precipitate in the gel, hindering transfer. Adding SDS to a final concentration of 0.1% in the transfer buffer will discourage this. Methanol tends to remove SDS from proteins, so reducing the methanol percentage to 10% or less will also guard against precipitation. Lowering methanol in the transfer buffer also promotes swelling of the gel, allowing large proteins to transfer more easily. Methanol is only necessary if using nitrocellulose. If using PVDF, methanol can be removed from the transfer buffer altogether, and is only needed to activate the PVDF before assembling the gel/membrane sandwich. Choose wet transfer overnight at 4°C instead of semi-dry transfer. The following reference discusses a gel and buffer system that allows transfer of proteins as large as 500 kD: Bolt and Mahoney, High-efficiency blotting of proteins of diverse sizes following sodium dodecyl sulfate–polyacrylamide, gel electrophoresis. Analytical Biochemistry 247, 185–192 (1997). http://(https://www.abcam.com/index.html?pageconfig=resource&rid=11404)

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Answer

The doublet when using serum may be result of mild proteolysis or incomplete reduction. I would recommend making sure protease inhibitors are added to your lysis buffer directly before use and heat your samples in laemmli buffer for 20 minutes at 72C.

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Answer

We have issued a free of charge replacement.

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Answer

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Answer

Vielen Dank für diese weiteren Informationen.
Es tut mir Leid, dass Sie Probleme mit diesem Antikörper hatten. Wie am Telefon besprochen, habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 123456 und sollte morgen bei Ihnen ankommen.
Ich kann die Blots bezüglich der KO Mäuse gut gebrauchen: Das gibt mir den Rückhalt gegenüber dem Labor nicht nur das betroffene Lot zurück zurufen, sondern dass die Lots in Produktion noch einmal genauer angeschaut werden!
Ich wünsche Ihnen viel Erfolg mit Ihren Projekten. Bitte melden Sie sich wieder bei uns, falls es weitere Fragen oder Probleme gibt.

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Answer

Unfortunately the discount code is only applicable before purchasing the antibodies to be tested. The discount code is not applicable neither for testing the isotype control as it has already been tested with Flow Cytometry, and therefore is not eligible for our antibody evaluation program (Abreview). However, you can still send us the results obtained from testing both primaries with Flow Cytometry and submit them in the form of an Abreview and earn 120 Abpoints exchangeable for rewards. If an image is submitted as well you’ll get 100 extra Abpoints. To find out more about our Abpoints, please refer to the next link: https://www.abcam.com/index.html?pageconfig=loyalty_about&viapagetrap=abpoints Just for your information I would like you to encourage having a look at our Flow Cytometry protocols online, where you could find useful information about the Flow Cytometry protocol as well as helpful tips: https://www.abcam.com/index.html?pageconfig=popular_protocols I hope this information was helpful. Please do not hesitate to contact us if you need further advice or information.

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Answer

We have not tested this specifically. I would assume it detects both but not familiar with the research of how you would be able to distinguish the plasma and cellular forms (by cell type?).

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Answer

Thank you for your enquiry. I would recommend that you aliquot your antibodies and only dilute when you are ready to use the antibody. Also, I would recommend that you dilute them into PBS, not TBS. Unfortunately, we do not have a protocol specifically for dilutions. Do be careful about how much you aliquot though, because you want to avoid doing a lot of freezing and thawing of the same aliquot. So try to aliquot enough for one use. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your enquiry. First, let me direct you a very informative website about IHC called IHC World. The web address is www.ihcworld.com. You can search that website for streptavidin, and then click on the Immunohistochemistry Introduction link. From there you can read about the different methods including the ABC and LSAB(involves streptavidin) methods. Here is what the site says about the streptavidin method: Streptavidin, derived from streptococcus avidini, is a recent innovation for substitution of avidin. The streptavidin molecule is uncharged relative to animal tissue, unlike avidin which has an isoelectric point of 10, and therefore electrostatic binding to tissue is eliminated. In addition, streptavidin does not contain carbohydrate groups which might bind to tissue lectins, resulting in some background staining. LSAB is technically similar to standard ABC method. The first layer is unlabeled primary antibody. The second layer is biotinylated secondary antibody. The third layer is Enzyme-Streptavidin conjugates (HRP-Streptavidin or AP-Streptavidin) to replace the complex of avidin-biotin peroxidase. The enzyme is then visualized by application of the substrate chromogen solutions to produce different colorimetric end products. The third layer can also be Fluorescent dye-Streptavidin such as FITC-Streptavidin if fluorescence labeling is preferred. A recent report suggests that LSAB method is about 5 to 10 times more sensitive than standard ABC method. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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