Recombinant Anti-Fibronectin antibody [F1] - Low endotoxin, Azide free (ab219366)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [F1] to Fibronectin - Low endotoxin, Azide free
- Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)
- Reacts with: Human
Overview
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Product name
Anti-Fibronectin antibody [F1] - Low endotoxin, Azide free
See all Fibronectin primary antibodies -
Description
Rabbit monoclonal [F1] to Fibronectin - Low endotoxin, Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Human serum and stomach tissue.This antibody gave a positive result in IF/ICC when used in the following formaldehyde fixed cell lines: HepG2. Flow Cyt (intra): HepG2
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General notes
ab219366 is the carrier-free version of ab32419.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
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.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
F1 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab219366 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 |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 263 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 263 kDa. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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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
modificationsSulfated.
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
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Database links
- Entrez Gene: 2335 Human
- Omim: 135600 Human
- SwissProt: P02751 Human
- Unigene: 203717 Human
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Alternative names
- CIG antibody
- Cold insoluble globulin antibody
- Cold-insoluble globulin antibody
see all
Images
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Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab32419 at a dilution of 1/250. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32419).
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Clone F1 (ab219366) has been successfully conjugated by Abcam. This image was generated using Anti-Fibronectin antibody [F1] (Alexa Fluor® 488). Please refer to ab198933 for protocol details.
ab198933 staining Fibronectin in A431 (Human epidermoid carcinoma cell line) cells. The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab198933 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Clone F1 (ab219366) has been successfully conjugated by Abcam. This image was generated using Anti-Fibronectin antibody [F1] (Alexa Fluor® 647). Please refer to ab198934 for protocol details.
ab198934 staining Fibronectin in HepG2 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab198934 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HepG2 cells fixed with 100% methanol (5min).
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Intracellular Flow Cytometry analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Fibronectin with purified ab32419 at 1/20 dilution (10 µg/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32419).
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ICC/IF image of unpurified ab32419 stained human mesenchymal stem cells. The cells were fixed in paraformaldehyde and then incubated in 0.1%BSA / 1% goat serum for 30 minutes, to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32419, 1/100 dilution) for 2 hours at 22°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG. DAPI was used to stain the cell nuclei (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32419).
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ICC/IF image of unpurified ab32419 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 ab32419 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32419).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (10)
ab219366 has been referenced in 10 publications.
- Qin S et al. Fibronectin protects lung cancer cells against docetaxel-induced apoptosis by promoting Src and caspase-8 phosphorylation. Tumour Biol 37:13509-13520 (2016). Human . PubMed: 27465556
- Cai JQ et al. Upregulation of HOXB7 promotes the tumorigenesis and progression of gastric cancer and correlates with clinical characteristics. Tumour Biol N/A:N/A (2015). PubMed: 26307396
- Ji X et al. The Anti-fibrotic Effects and Mechanisms of MicroRNA-486-5p in Pulmonary Fibrosis. Sci Rep 5:14131 (2015). PubMed: 26370615
- Nutter F et al. Different molecular profiles are associated with breast cancer cell homing compared with colonisation of bone: evidence using a novel bone-seeking cell line. Endocr Relat Cancer 21:327-41 (2014). PubMed: 24413608
- Berardis S et al. Gene expression profiling and secretome analysis differentiate adult-derived human liver stem/progenitor cells and human hepatic stellate cells. PLoS One 9:e86137 (2014). IHC-P ; Human . PubMed: 24516514
- Gerdes MJ et al. Highly multiplexed single-cell analysis of formalin-fixed, paraffin-embedded cancer tissue. Proc Natl Acad Sci U S A 110:11982-7 (2013). ICC/IF ; Human . PubMed: 23818604
- Wakahashi S et al. VAV1 represses E-cadherin expression through the transactivation of Snail and Slug: a potential mechanism for aberrant epithelial to mesenchymal transition in human epithelial ovarian cancer. Transl Res 162:181-90 (2013). WB ; Human . PubMed: 23856093
- Hubbard B et al. Heparin-dependent regulation of fibronectin matrix conformation. Matrix Biol N/A:N/A (2013). PubMed: 24148804
- Sun Q et al. Role of myocyte enhancing factor 2B in epithelial myofibroblast transition of human gingival keratinocytes. Exp Biol Med (Maywood) 237:178-85 (2012). PubMed: 22302709
- Mirabet V et al. Long-term storage in liquid nitrogen does not affect cell viability in cardiac valve allografts. Cryobiology 57:113-21 (2008). IHC-Fr . PubMed: 18703039