Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [F14] to Fibronectin
- Suitable for: IHC-Fr, WB, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-Fibronectin antibody [F14]
See all Fibronectin primary antibodies
DescriptionRabbit monoclonal [F14] to Fibronectin
Tested applicationsSuitable for: IHC-Fr, WB, ICC/IF, Flow Cytmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Mouse, Rat, Human
Recombinant full length protein corresponding to Human Fibronectin aa 1-2400.
Database link: P02751
- WB: Human, mouse and rat serum lysate. IHC-Fr: Rat kidney tissue. ICC/IF: HepG2 cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.21% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab45688 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|WB||1/1000 - 1/10000. Detects a band of approximately 263 kDa (predicted molecular weight: 263 kDa).|
For unpurified use at 1/250.
FunctionFibronectins 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 specificityPlasma 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 diseaseGlomerulopathy with fibronectin deposits 2
Sequence similaritiesContains 12 fibronectin type-I domains.
Contains 2 fibronectin type-II domains.
Contains 16 fibronectin type-III domains.
Developmental stageUgl-Y1, Ugl-Y2 and Ugl-Y3 are present in the urine from 0 to 17 years of age.
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 localizationSecreted, extracellular space, extracellular matrix.
- Information by UniProt
- CIG antibody
- Cold insoluble globulin antibody
- Cold-insoluble globulin antibody
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Fibronectin knockout HAP1 whole cell lysate (20 µg)
Lanes 1 - 2: Merged signal (red and green). Green - ab45688 observed at 262 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab45688 was shown to react with Fibronectin in wild-type HAP1 cells as signal was lost in Fibronectin knockout cells. Wild-type and Fibronectin knockout samples were subjected to SDS-PAGE. Ab45688 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/2000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Fibronectin with purified ab45688 at 1:150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Fibronectin with Purified ab45688 at 1:500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Anti-Fibronectin antibody [F14] (ab45688) at 1/10000 dilution (purified) + Human serum lysates at 15 µg
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 263 kDa
Blocking and diluting buffer: 5% NFDM/TBST.
Unpurified ab45688 staining Fibronectin in rat kidney tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with Cas-Block for 30 minutes at 25°C. Samples were incubated with primary antibody (1/250) for 16 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.
All lanes : Anti-Fibronectin antibody [F14] (ab45688) at 1/10000 dilution (purified)
Lane 1 : Mouse serum lysates
Lane 2 : Rat serum lysates
Lysates/proteins at 15 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 263 kDa
Observed band size: 263 kDa
Blocking and diluting buffer: 5% NFDM/TBST
Anti-Fibronectin antibody [F14] (ab45688) at 1/5000 dilution (unpurified) + human serum
Predicted band size: 263 kDa
Observed band size: 263 kDa
ICC/IF image of unpurified ab45688 stained HepG2 (Human liver hepatocellular carcinoma cell line) cells. The cells were fixed in 4% formaldehyde (10 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab45688 at 1/250 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab45688 has been referenced in 35 publications.
- He J et al. Kidney targeted delivery of asiatic acid using a FITC labeled renal tubular-targeting peptide modified PLGA-PEG system. Int J Pharm 584:119455 (2020). PubMed: 32464235
- Pang X et al. Hirudin Reduces the Expression of Markers of the Extracellular Matrix in Renal Tubular Epithelial Cells in a Rat Model of Diabetic Kidney Disease Through the Hypoxia-Inducible Factor-1a (HIF-1a)/Vascular Endothelial Growth Factor (VEGF) Signaling Pathway. Med Sci Monit 26:e921894 (2020). PubMed: 32473006
- Lian H et al. Malignant fibrous histiocytoma amplified sequence 1 alleviates inflammation and renal fibrosis in diabetic nephropathy by inhibiting TLR4. Biosci Rep 39:N/A (2019). PubMed: 31696221
- Li Y et al. M10 peptide attenuates silica-induced pulmonary fibrosis by inhibiting Smad2 phosphorylation. Toxicol Appl Pharmacol 376:46-57 (2019). PubMed: 31125577
- Pan J et al. Pterostilbene, a bioactive component of blueberries, alleviates renal fibrosis in a severe mouse model of hyperuricemic nephropathy. Biomed Pharmacother 109:1802-1808 (2019). PubMed: 30551434