Recombinant Anti-Fibronectin antibody [EPR23110-46] KO Tested (ab268020)

Product name

Anti-Fibronectin antibody [EPR23110-46]
See all Fibronectin primary antibodies
Description

Rabbit monoclonal [EPR23110-46] to Fibronectin
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. ICC/IF: HepG2 and NIH/3T3 cells.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23110-46
Isotype

IgG
Research areas

Alternative Versions
- Anti-Fibronectin antibody [EPR23110-46] - BSA and Azide free (ab268022)
Compatible Secondaries
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Related Products
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab268020 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		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF		1/50.
WB		1/1000. Detects a band of approximately 285 kDa (predicted molecular weight: 262 kDa). This antibody can be used at 0.585 μg/ml.

Notes
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF 1/50.
WB 1/1000. Detects a band of approximately 285 kDa (predicted molecular weight: 262 kDa). This antibody can be used at 0.585 μg/ml.

Application notes

Is unsuitable for Flow Cyt or IP.

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.
Target information above from: UniProt accession P02751 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2335 Human
- Entrez Gene: 14268 Mouse
- Entrez Gene: 25661 Rat
- Omim: 135600 Human
- SwissProt: P02751 Human
- SwissProt: P11276 Mouse
- SwissProt: P04937 Rat
- Unigene: 203717 Human
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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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).
Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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.
Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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).
Western blot - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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?

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.
Western blot - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

All lanes : Anti-Fibronectin antibody [EPR23110-46] (ab268020) at 1/1000 dilution

Lane 1 : Human serum
Lane 2 : Human plasma
Lane 3 : Human kidney tissue lysate
Lane 4 : Human stomach tissue lysate
Lane 5 : Mouse plasma
Lane 6 : Rat plasma

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 5-6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 262 kDa
Observed band size: 258 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-2, 5-6: 3 seconds; Lane 3: 125 seconds; Lane 4: 33 seconds.

The expression molecular weight observed is consistent with what has been described in the literature (PMID:28367057).
Immunohistochemistry (Frozen sections) - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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).
Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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.
Anti-Fibronectin antibody [EPR23110-46] (ab268020)

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

SDS download

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Publishing research using ab268020? Please let us know so that we can cite the reference in this datasheet.

ab268020 has not yet been referenced specifically in any publications.

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Developmental stage

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications