MA5-11981

antibody from Invitrogen Antibodies

Targeting: FN1 CIG, FINC, GFND2, LETS, MSF

Provider product page for MA5-11981

Western blot

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [34]
Comments [0]
Validations
Western blot [3]
Immunohistochemistry [2]
Other assay [17]

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Product number: MA5-11981 - Provider product page
Provider: Invitrogen Antibodies
Product name: Fibronectin Monoclonal Antibody (FBN11)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-11981 targets Fibronectin in IF, WB and IHC (P) applications and shows reactivity with Human, mouse, and Rat samples.
Antibody clone number: FBN11
Concentration: 0.2 mg/mL

FN1 protein structure - MA5-11981 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of fibronectin was performed by loading insoluble fibronectin isolated via a deoxycholate assay from 3 x 105 mouse smooth muscle cells from three wild type (WT) or integrin knock-out (KO) mice per well onto an SDS-PAGE gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with a fibronectin monoclonal antibody (Product # MA5-11981) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:2000 for 2 hours. Detection was performed using SuperSignal West Pico (Product # 34080). Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockout of Fibronectin was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR616644_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Fibronectin was performed by loading 30 µg of Hep G2 Wild Type (Lane 1), Hep G2 Wild Type treated with 1X PTI for 4hrs (Lane 2), Hep G2 Cas9 (Lane 3), Hep G2 Cas9 treated with 1X PTI for 4hrs (Lane 4), Hep G2 Fibronectin KO (Lane 5) and Hep G2 Fibronectin KO treated with 1X PTI for 4hrs (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Fibronectin Monoclonal Antibody (FBN11) (Product # MA5-11981, 1:2,000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Fibronectin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Fibronectin Monoclonal Antibody (FBN11) (Product # MA5-11981) and a ~260kDa band corresponding to Fibronectin was observed across cell lines tested . Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with 1X PTI for 4h (Lane 2), Hep G2 treated with 10uM FLI-06 for 4h (Lane 3), HEL 92.1.7 (Lane 4), HEL 92.1.7 treated with 1X PTI for 4h (Lane 5), HEL 92.1.7 treated with 10uM FLI-06 for 4h (Lane 6) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Anti-Fibronectin Monoclonal Antibody (FBN11) (Product # MA5-11981) showed enhanced pick up in positive cell line Hep G2 upon treatment with secretion blockers as compared to in negative cell line HEL 92.1.7 treated with the same blockers.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Gallic acid pretreatment suppressed cardiac fibrosis in isoproterenol (ISP)-treated mice. Gallic acid administration was started 2 weeks before infusion of ISP (3 days) in mice; sham + vehicle, ISP + vehicle, and ISP + gallic acid group. ( A ) Representative images of hearts stained with Masson's trichrome, which were used to examine fibrosis (collagen deposition, scale bar, 100 mum). ( B ) Immunofluorescent staining with anti-collagen type I of heart sections from ISP-induced mice (scale bar, 50 mum). ( C ) Protein lysates from heart tissues were subjected to western blot with anti-collagen type I, anti-fibronectin, and anti-alpha-smooth muscle actin (SMA) antibodies. GAPDH was used as a loading control. ( D ) Collagen type I and fibronectin proteins were quantified by densitometry. ( E - G ) The mRNA levels of fibrosis-related genes (collagen type I, fibronectin, SMA) were analyzed by qRT-PCR. Data are presented as the means +- SD of 6~15 mice per group. * P < 0.05, * P < 0.01, and *** P < 0.001 versus sham + vehicle; ## P < 0.01versus ISP + vehicle.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 SHED-derived SMCs promote EC vessel formation. a Photomicrographs of EC-coated microbeads cultured in fibrin gels with SMC, SHED and SHED-induced SMC on the gel surface. b The schematic representation of calculating tube formation from each bead: (i) the number of vessels which sprout from the bead directly (in blue ); (ii) the total number of individual vessel segments (including blue and yellow ); and (iii) the total length of all the vessel segments. c Quantification of the vessel structures in the fibrin gel bead assay in the presence of SMC, SHED and SHED-induced SMC.*: p < 0.05 versus SMCs group, ^: p < 0.05 between SHED and SHED-derived SMCs. d Western blot showing fibronectin expression within each experimental group. All experiments were performed three times ( N = 3). ECs endothelial cells, SHED stem cells from human exfoliated deciduous teeth, SMCs smooth muscle cells

Supportive validation

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Experimental details: 5 FIGURE Lysophosphatidic acid (LPA)-induced neutrophil extracellular traps (NETs) bound plasma proteins important for thrombus stability. Neutrophils without stimulation and NETs induced by LPA (10 muM, 240 min) were seeded on glass coverslips and incubated with 50% plasma in phosphate buffered saline. Samples were stained for (A) von Willebrand factor, (B) fibrinogen, and (C) fibronectin (400x) (n = 4)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 TSA regulates TGF -beta1-mediated ECM proteins and E-cadherin in HK 2 cells. ( A ) HK2 cells were treated with increasing concentrations of TSA (0, 10, 25, 50 100 and 250 nM) for 24 hrs and cell viability was measured by the MTT assay. ( B ) HK2 cells were co-treated with TGF-beta1 (5 ng/ml) and TSA (100 nM). ECM proteins and EMT markers were determined using Western blot analysis. Representative immunoblots are shown. beta-Actin was used as the loading control. ( C-F ) Quantification was performed by densitometry for at least four independent experiments. ( G ) Representative immunofluorescence images for E-cadherin in HK2 cells. Immunofluorescence staining was performed using anti-E-cadherin antibody and DAPI was used for staining cell nuclei. Merged images are shown. Scale bar represents 50 mum. *** P < 0.001 compared with untreated cells. # P < 0.05 and ### P < 0.001 compared with TGF-beta1-treated cells. NS, not significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 PCI 34051 did not affect TGF -beta1-mediated EMT markers in HK 2 cells. ( A ) Cytotoxicity of PCI 34051 in HK 2 cells. ( B ) HK 2 cells were co-treated with TGF -beta1 (5 ng/ml) and PCI 34051 (5 muM) for 24 hrs. Western blot analysis was performed using collagen type I (collagen 1), fibronectin, N-cadherin, and E-cadherin antibodies. beta-Actin was used as the loading control. ( C-F ) Quantification was performed by densitometry for at least four independent experiments. ** P < 0.01 and *** P < 0.001 compared with untreated cells. # P < 0.05 and ### P < 0.001 compared with TGF -beta1 treated cells. NS , not significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 LMK 235 did not affect TGF -beta1-mediated ECM proteins and EMT markers in HK 2 cells. ( A ) Cytotoxicity of LMK 235 in HK 2 cells. ( B ) HK 2 cells were co-treated with TGF -beta1 and LMK 235 for 24 hrs. Western blot analysis was performed using collagen I, fibronectin, N-cadherin, and E-cadherin antibodies. beta-Actin was used as the loading control. ( C-F ) Quantification was performed by densitometry for at least four independent experiments. * P < 0.05 and *** P < 0.001 compared with untreated cells. ## P < 0.01 and ### P < 0.001 compared with TGF -b1 treated cells. NS , not significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 TGF-beta-induced cell migration and invasion was reversed by knockdown of TAGLN . ( A ) Cell migration and invasion in HCT116 cells (left panel) and representative images of crystal violet staining (right panel). ( B ) Cell migration and invasion in HT29 cells (left panel) and representative images of crystal violet staining (right panel). ( C ) Representative images of Western blotting assay of E-cadherin, vimentin, fibronectin, MMP2, MMP9, and GADPH . 200 x magnification. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Overexpression of HMGA2 restored altered CRC cell migration and invasion induced by inhibition of TAGLN . ( A ) Cell migration and invasion in HCT116 cells (left panel) and representative images of crystal violet staining (right panel). ( B ) Cell migration and invasion in HT29 cells (left panel) and representative images of crystal violet staining (right panel). ( C ) Representative images of Western blotting assay of nucleus and total TAGLN levels. ( D ) Representative images of Western blotting assay of E-cadherin, vimentin, fibronectin, MMP2, MMP9, and GADPH . 200 x magnification. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 TGF-beta-induced TAGLN expression increased migration and invasion of tumor promoting. ( A ) Image of tumors in nude mice inoculated with HT29 cells (Blank) and HT29 cells pretreated with TGF-beta and si- TAGLN (TGF-beta+si- TAGLN group). ( B ) Tumor volume was measured every four days. Tumor volume=1/2*L*W*W. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIG 6 Inhibition of Fn fibril formation increases the bacterial uptake in osteoblasts. Host cells were cultured for 2 days with a 70-kDa fragment of fibronectin to block Fn assembly. (A) Representative immunofluorescence microscopy images of Fn expression on pHOBs detected by anti-Fn antibody. (B) A549 cells and pHOBs cultured with the 70-kDa fragment of Fn as indicated and infected with S. aureus 6850 or S. carnosus TM300(pFnBA4) (MOI 50). One hour post infection, extracellular staphylococci were removed by lysostaphin treatment. Subsequently, the host cells were detached and the number of host cells was determined. Host cells were lysed and the number of viable intracellular bacteria was assessed by plate counting. Numbers of intracellular bacteria in non-pretreated cells were set to 100%. Data are means +- SD from 3 independent experiments. *, P < 0.05, one-way ANOVA followed by Dunnett's multiple-comparison test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 PCI34051 alleviates cardiac fibrosis in transverse aortic constriction (TAC) mice through the TGF- beta 1-Smad2/3 pathway. (a, b) Picrosirius red staining of mouse cardiac tissues; representative images and quantification are shown. Scale bar = 50 mu m. The expression levels of the fibrosis marker genes Col1a1 (c), Fn1 (d), Acta2 (e), and Tgfb1 (f) were determined using quantitative real-time polymerase chain reaction ( n = 5-6 per group). (g) The cardiac expression levels of Fn1, Acta2, Tgfb1, p-Smad2/3, and Smad2/3 were analyzed using western blotting. Actb was used as a loading control. Representative blots are shown. (h-k) Quantification of Fn1, Acta2, Tgfb1, and p-Smad2/3 to Smad2/3 ( n = 5-6 per group).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 PCI34051 alleviates pulmonary congestion and fibrosis in transverse aortic constriction (TAC) mice. (a) The ratio of lung weight-to-bodyweight (LW/BW) in the sham+vehicle, sham+PCI34051 (10 mg/kg bodyweight/day), TAC+vehicle, and TAC+PCI34051 (3, 10, or 30 mg/kg bodyweight/day) groups ( n = 5-6 per group) was determined. Representative images of mouse lungs (B). (b) Representative images of pulmonary tissues stained with hematoxylin and eosin. Scale bar = 50 mu m. (c) Representative images of pulmonary tissues stained with Picrosirius red. Scale bar = 50 mu m. (d) Pulmonary fibrosis was quantified using ImageJ software. (e-h) The mRNA levels of Col1a1 , Fn1 , Acta2 , and Tgfb1 were evaluated using quantitative real-time polymerase chain reaction ( n = 5-6 per group). (i-m) Representative western blots and quantification of Fn1, Acta2, Tgfb1, p-Smad2/3, and Smad2/3 levels in the pulmonary tissues ( n = 5-6 per group).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 PCI34051 or Hdac8 knockdown mitigates TGF- beta 1-induced fibrosis in rat cardiac fibroblasts. (a-e) Rat cardiac fibroblasts were pre-treated with TGF- beta 1 (10 ng/mL) for 1 h and cultured in the presence of vehicle or PCI34051 (10 mu M) for 8 h. mRNA levels of Hdac8 , Col1a1 , Fn1 , Acta2 , and Tgfb1 were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR) ( n = 4-6 per group). (f-k) Representative immunoblots and quantification of Hdac8, Fn1, Acta2, Tgfb1, p-Smad2/3, and Smad2/3 levels in rat cardiac fibroblasts ( n = 6 per group). Actb was used as a loading control. (l-p) Rat cardiac fibroblasts were transfected with control or short-interfering RNA against Hdac8 and treated with TGF- beta 1 (10 ng/mL) for 9 h. The mRNA levels of Hdac8 , Fn1 , Acta2 , Tgfb1 , and Ace1 were determined using qRT-PCR ( n = 5 per group). (q-w) Representative immunoblots and quantification of Hdac8, Ace1, Fn1, Acta2, Tgfb1, p-Smad2/3, and Smad2/3 levels in rat cardiac fibroblasts ( n = 4 per group). Actb was used as a loading control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Immunohistochemistry of agarose gels following: (A) 1 and 7 days of culture in standard medium at pH 7.4 and (B) a subsequent 24 hours at pH 7.1 or 6.5. Nucleus pulposus (NP) cells/agarose gels were snap frozen in liquid nitrogen and cryosectioned. Antibodies against the pericellular marker type VI collagen, the integrin receptor alpha5beta1, and separate subunits alpha5 and beta1 and the extracellular matrix protein fibronectin (the predominant ligand of alpha5beta1 integrins) were used to probe sections. (A) NP cells did not appear to express type VI collagen or alpha5beta1 integrin following 1 day of preculture. However, NP cells did express both type VI collagen and alpha5beta1 following 7 days of preculture (black arrows indicate cells expressing proteins which were not expressed at day 0), (B) with 24 hours of culture at pH 7.1 or 6.5 not affecting this expression. (A) NP cells did express the alpha5 and beta1 integrin subunits, as well as fibronectin, following 1 and 7 days of preculture in standard medium, (B) with culture at pH 7.1 or 6.5 having no noticeable affect on the expression

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 CGSA showing axolotl wound healing signatures in AGMs. a Heatmap of Wound Healing Network genes in axolotl wound repair and acute SIV infection, ordered by day 1 of aquatic axolotl wound repair. The biological functions that correlate with D1 axolotl wound repair, and their associated genes are shown on the right. b A protein interaction network between proteins encoded by genes in the Wound Healing Network. Red and blue nodes indicate up and downregulation in aquatic (left) and terrestrial (right) axolotl at day 1. c Immunohistochemistry staining of fibronectin (FN) in rectal mucosa at baseline, D1 and D3 post SIV inoculation. Right panels show quantification by pixel density, middle and left panels show fibronectin protein expression in lamina propria. Representative images are shown below. The scale bar included in each image represents a length of 50 mum