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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

Reviews (3) Submit a question
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Western blot - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
  • Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23110-46] to Fibronectin - BSA and Azide free
  • Suitable for: IHC-P, IHC-Fr, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 568 Unconjugated

Overview

  • Product name

    Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free
    See all Fibronectin primary antibodies

  • Description

    Rabbit monoclonal [EPR23110-46] to Fibronectin - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IHC-Fr, ICC/IF, WBmore details
    Unsuitable for: Flow Cyt or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human serum, kidney and stomach lysate. Mouse, human and rat plasma lysate. NIH/3T3 whole cell lysate; HAP1 whole cell lysate. IHC-P: Human mammary gland tissue. Human esophageal squamous cell carcinoma tissue. Mouse and rat stomach tissue, human pancreatic carcinoma tissue. ICC/IF: HepG2 and NIH/3T3 cells.

  • General notes

    ab268022 is the carrier-free version of ab268020.

    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 compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab268022 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
IHC-P (1)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr (1)
Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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).
Notes
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.
IHC-Fr
Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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).
Application notes
Is unsuitable for Flow Cyt or IP.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of paraffin-embedded Human pancreatic carcinoma tissue labeling Fibronectin with ab268020 at 1/4000 dilution (0.146 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). The section was incubated with ab268020 at 4°C overnight. Positive staining in the human pancreatic carcinoma counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP)(ab214880)

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Fibronectin with ab268020 at 1/2000 dilution (0.29 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the smooth muscle of rat stomach (PMID:650101) Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Fibronectin with ab268020 at 1/2000 dilution (0.29 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the smooth muscle of mouse stomach (PMID:650101) Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of paraffin-embedded human esophageal squamous cell carcinoma tissue labeling Fibronectin with ab268020 at 1/2000 dilution (0.29 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the stroma of human esophageal squamous cell carcinoma (PMID:30314454). Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling Fibronectin with ab268020 at 1/2000 dilution (0.29 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in the stroma of human mammary gland (PMID:650101). Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat colon tissue labeling Fibronectin with ab268020 at 1/100 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining mainly on smooth muscle of rat colon (PMID: 17881565) is observed. The nuclear counterstain is DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse colon tissue labeling Fibronectin with ab268020 at 1/100 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Positive staining mainly on smooth muscle of mouse colon (PMID: 17881565) is observed. The nuclear counterstain is DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Western blot - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Western blot - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      All lanes : Anti-Fibronectin antibody [EPR23110-46] (ab268020) at 0.585 µg/ml (1:1000 dilution)

      Lane 1 : Human stomach tissue lysate at 10 µg
      Lane 2 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg
      Lane 3 : Fibronectin-1 knockout HAP1 whole cell lysate at 40 µg
      Lane 4 : Wild-type HAP1 whole cell lysate at 40 µg

      Developed using the ECL technique.

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



      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

      ab268020 was shown to specifically 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. ab268020 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      Exposure time: Lanes 1-2: 37 seconds; Lanes 3-4: 3 minutes.

      Blocking/diluting buffer and concentration: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Fibronectin with ab268020 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cells is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling Fibronectin with ab268020 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cells is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab268020).

    • Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
      Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)

    Protocols

    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.

    Click here to view the general protocols

    References (0)

    Publishing research using ab268022? Please let us know so that we can cite the reference in this datasheet.

    ab268022 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris EDTA pH=9
    Permeabilization
    Yes - Triton 0.1%
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Olivia Lenoir

    Verified customer

    Submitted Mar 06 2023

    Immunohistochemistry (Frozen sections) abreview for Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Glioblastoma multiforme)
    Permeabilization
    Yes - 0.25% TritonX-100
    Specification
    Glioblastoma multiforme
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 4%
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 24 2022

    Mass Cytometry abreview for Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free

     Excellent
    Abreviews
    abreview image
    Application
    Mass Cytometry
    Sample
    Mouse Tissue sections (Mouse Lung)
    Specification
    Mouse Lung
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Tara Sutherland

    Verified customer

    Submitted Feb 19 2021

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