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Fibronectin Recombinant Rabbit Monoclonal Antibody [JF0582]

Usd: 385

Specification

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Catalog# ET1702-25

Fibronectin Recombinant Rabbit Monoclonal Antibody [JF0582]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • mIHC

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Fibronectin Recombinant Rabbit Monoclonal Antibody [JF0582]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant full length protein of Human Fibronectin aa 1-2477 / 2477.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, mIHC

Molecular Weight

Predicted band size: 272 kDa

Positive Control

HepG2 cell lysate, Caco-2 cell lysate, SK-MEL-28 cell lysate, human kidney tissue lysate, MEF cell lysate, NIH/3T3 cell lysate, MG-63 cell lysates, HepG2, Hela, NIH/3T3, mouse liver tissue.

Conjugation

unconjugated

Clone Number

JF0582

RRID

AB_3070288

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100-1:500

  • IF-Tissue

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • mIHC

  • 1:100

Target

Function

Fibronectin is an extracellular matrix glycoprotein present on most cell surfaces, in extracellular fluids and in plasma. A high molecular weight heterodimeric protein, it was originally discovered as a protein missing from the surfaces of virus-transformed cells, and it has been shown to be involved in various functions including cell adhesion, cell motility and wound healing. Alternative splicing and glycosylation give rise to several different forms of Fibronectin, some of which exhibit restricted tissue distribution or association with malignancies. It has been shown that Myofibroblast phenotype formation correlates with the occurrence of glycosylated Fibronectin and Fibronectin splice variants in Dupuytren's disease.

Background References

1. Patten J et al. Fibronectin in development and wound healing. Adv Drug Deliv Rev. 2021 Mar

2. Dalton CJ et al. Fibronectin: Molecular Structure, Fibrillar Structure and Mechanochemical Signaling. Cells. 2021 Sep

Tissue Specificity

Expressed in the inner limiting membrane and around blood vessels in the retina (at protein level). 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.

Post-translational Modification

Sulfated.; It is not known whether both or only one of Thr-2155 and Thr-2156 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.; Some lysine residues are oxidized to allysine by LOXL3, promoting fibronectin activation and matrix formation.

Subcellular Location

Secreted, extracellular space, extracellular matrix.

UNIPROT

SwissProt: P02751 Human

SwissProt: P11276 Mouse

Synonyms

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

Expand

Images

  • Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: Caco-2 cell lysate<br />Lane 3: SK-MEL-28 cell lysate<br />Lane 4: Human kidney tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 272 kDa<br />Observed band size: 272 kDa<br /><br />Exposure time: 3 minutes 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (ET1702-25) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: Caco-2 cell lysate
    Lane 3: SK-MEL-28 cell lysate
    Lane 4: Human kidney tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 272 kDa
    Observed band size: 272 kDa

    Exposure time: 3 minutes 20 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-25) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: MEF cell lysate<br />Lane 3: NIH/3T3 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 272 kDa<br />Observed band size: 272 kDa<br /><br />Exposure time: 2 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (ET1702-25) at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: MEF cell lysate
    Lane 3: NIH/3T3 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 272 kDa
    Observed band size: 272 kDa

    Exposure time: 2 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-25) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Fibronectin on MG-63 cell lysates with Rabbit anti-Fibronectin antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 272 kDa<br />Observed band size: 272 kDa<br /><br />Exposure time: 2 minutes;<br /><br />6% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Fibronectin on MG-63 cell lysates with Rabbit anti-Fibronectin antibody (ET1702-25) at 1/1,000 dilution.

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 272 kDa
    Observed band size: 272 kDa

    Exposure time: 2 minutes;

    6% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-25) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • ICC staining of Fibronectin in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Fibronectin in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-25, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of Fibronectin in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Fibronectin in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-25, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of Fibronectin in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Fibronectin in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-25, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Fibronectin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Fibronectin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-25, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2-si NT cell lysate<br />Lane 2: HepG2-si Fibronectin cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 272 kDa<br />Observed band size: 272 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-25" style="font-weight: bold;text-decoration: underline;">ET1702-25</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (ET1702-25) at 1/2,000 dilution.

    Lane 1: HepG2-si NT cell lysate
    Lane 2: HepG2-si Fibronectin cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 272 kDa
    Observed band size: 272 kDa

    Exposure time: 20 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-25) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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