Recombinant Anti-FOXC1 antibody [EPR20685] KO Tested (ab227977)

Product name

Anti-FOXC1 antibody [EPR20685]
See all FOXC1 primary antibodies
Description

Rabbit monoclonal [EPR20685] to FOXC1
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, Flow Cyt (Intra), IP, IHC-P, IHC-Fr, WBmore details
Species reactivity

Reacts with: Mouse, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: MDA-MB-231, HEK-293T and HeLa whole cell lysates; Human fetal kidney lysate. IHC-P: Human gastric cancer and basal-like breast cancer tissues; Mouse cerebrum tissue. IHC-Fr: Mouse fetal brain E14.5 tissue. ICC/IF: HeLa and HEK-293T cells (HEK293-FOXC1 KO cells used as a negative control). Flow Cyt (intra): HEK-293T cells. IP: HeLa whole cell lysate.
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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20685
Isotype

IgG
Research areas

Alternative Versions
- Anti-FOXC1 antibody [EPR20685] - BSA and Azide free (ab230159)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab227977 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
ICC/IF		Use a concentration of 12 µg/ml.
Flow Cyt (Intra)		1/100.
IP		1/50.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		1/1000. Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
WB		1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 57 kDa). WB works for some mouse lysates.

Notes
ICC/IF Use a concentration of 12 µg/ml.
Flow Cyt (Intra) 1/100.
IP 1/50.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/1000. Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
WB 1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 57 kDa). WB works for some mouse lysates.

Function

Binding of FREAC-3 and FREAC-4 to their cognate sites results in bending of the DNA at an angle of 80-90 degrees.
Tissue specificity

Expressed in all tissues and cell lines examined.
Involvement in disease

Defects in FOXC1 are the cause of Axenfeld-Rieger syndrome type 3 (RIEG3) [MIM:602482]; also known as Axenfeld-Rieger syndrome (ARS) or Axenfeld syndrome or Axenfeld anomaly. It is characterized by posterior corneal embryotoxon, prominent Schwalbe line and iris adhesion to the Schwalbe line. Other features may be hypertelorism (wide spacing of the eyes), hypoplasia of the malar bones, congenital absence of some teeth and mental retardation. When associated with tooth anomalies, the disorder is known as Rieger syndrome. Glaucoma is a progressive blinding condition that occurs in approximately half of patients with Axenfeld-Rieger malformations.
Defects in FOXC1 are the cause of iridogoniodysgenesis anomaly (IGDA) [MIM:601631]. IGDA is an autosomal dominant phenotype characterized by iris hypoplasia, goniodysgenesis, and juvenile glaucoma.
Defects in FOXC1 are a cause of Peters anomaly (PAN) [MIM:604229]. Peters anomaly consists of a central corneal leukoma, absence of the posterior corneal stroma and Descemet membrane, and a variable degree of iris and lenticular attachments to the central aspect of the posterior cornea.
Sequence similarities

Contains 1 fork-head DNA-binding domain.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q12948 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2296 Human
- Entrez Gene: 17300 Mouse
- GenBank: NP_001444 Human
- Omim: 601090 Human
- SwissProt: Q12948 Human
- SwissProt: Q61572 Mouse
- Unigene: 348883 Human
- Unigene: 12949 Mouse
Alternative names
- ARA antibody
- FKH L7 antibody
- FKHL 7 antibody
- FKHL7 antibody
- Forkhead (Drosophila) like 7 antibody
- Forkhead box C1 antibody
- Forkhead box protein C1 antibody
- Forkhead drosophila homolog like 7 antibody
- Forkhead like 7 antibody
- Forkhead related activator 3 antibody
- Forkhead related protein FKHL7 antibody
- Forkhead related transcription factor 3 antibody
- Forkhead-related protein FKHL7 antibody
- Forkhead-related transcription factor 3 antibody
- FOX C1 antibody
- FOXC 1 antibody
- Foxc1 antibody
- FOXC1_HUMAN antibody
- FREAC 3 antibody
- FREAC-3 antibody
- FREAC3 antibody
- IGDA antibody
- IHG 1 antibody
- IHG1 antibody
- IRID 1 antibody
- IRID1 antibody
- Iridogoniodysgenesis type 1 antibody
- Myeloid factor delta antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-FOXC1 antibody [EPR20685] (ab227977)

ab227977 staining FOXC1 in wild-type HEK293 cells (top panel) and FOXC1 knockout HEK293 cells (bottom panel). The cells were fixed with paraformaldehyde (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 with ab227977 at 12 μg/ml and ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western blot - Anti-FOXC1 antibody [EPR20685] (ab227977)

All lanes : Anti-FOXC1 antibody [EPR20685] (ab227977) at 1/1000 dilution

Lane 1 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4 : Human fetal kidney lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 57 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Exposure time : Lanes 1 & 4: 3 minutes; Lanes 2-3: 5 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular mass is consistent with what has been described in the literature (PMID: 27708239).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXC1 antibody [EPR20685] (ab227977)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling FOXC1 with ab227977 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human gastric cancer (PMID:24329718). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXC1 antibody [EPR20685] (ab227977)

Immunohistochemical analysis of paraffin-embedded human basal-like breast cancer tissue labeling FOXC1 with ab227977 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human basal-like breast cancer (PMID:27708239; PMID:20406990). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXC1 antibody [EPR20685] (ab227977)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling FOXC1 with ab227977 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in the pericytes and endothelium of blood vessels in mouse cerebrum is observed (PMID:25733312; PMID: 23862012). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Immunohistochemistry (Frozen sections) - Anti-FOXC1 antibody [EPR20685] (ab227977)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse fetal brain E14.5 tissue labeling FOXC1 with ab227977 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive nuclear staining localized in the meninges and adjacent cortex region on mouse fetal brain (PMID: 23862012).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-FOXC1 antibody [EPR20685] (ab227977)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling FOXC1 with ab227977 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HEK-293T cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-FOXC1 antibody [EPR20685] (ab227977)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling FOXC1 with ab227977 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HEK-293T cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-FOXC1 antibody [EPR20685] (ab227977)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line labeling FOXC1 with ab227977 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-FOXC1 antibody [EPR20685] (ab227977)

FOXC1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab227977 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab227977 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: HeLa whole cell lysate 10 µg (Input).

Lane 2: ab227977 IP in HeLa whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227977 in HeLa whole cell lysate.

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

Exposure time: 10 seconds.
Anti-FOXC1 antibody [EPR20685] (ab227977)

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

Datasheet download

Download

Publishing research using ab227977? Please let us know so that we can cite the reference in this datasheet.

ab227977 has been referenced in 8 publications.

Sun B et al. Regulation of host and virus genes by neuronal miR-138 favours herpes simplex virus 1 latency. Nat Microbiol 6:682-696 (2021). PubMed: 33558653
Grönroos P et al. Directed Differentiation of Human Pluripotent Stem Cells towards Corneal Endothelial-Like Cells under Defined Conditions. Cells 10:N/A (2021). PubMed: 33562615
He T et al. A novel SIRT6 activator ameliorates neuroinflammation and ischemic brain injury via EZH2/FOXC1 axis. Acta Pharm Sin B 11:708-726 (2021). PubMed: 33777677
Jiang J et al. FOXC1 Negatively Regulates DKK1 Expression to Promote Gastric Cancer Cell Proliferation Through Activation of Wnt Signaling Pathway. Front Cell Dev Biol 9:662624 (2021). PubMed: 33987183
Sun H et al. MCM3AP-AS1 promotes cisplatin resistance in gastric cancer cells via the miR-138/FOXC1 axis. Oncol Lett 21:211 (2021). PubMed: 33510812
Xia W et al. Overexpression of Foxc1 regenerates crushed rat facial nerves by promoting Schwann cells migration via the Wnt/ß-catenin signaling pathway. J Cell Physiol 235:9609-9622 (2020). PubMed: 32391604
Chen H et al. GPX8 is transcriptionally regulated by FOXC1 and promotes the growth of gastric cancer cells through activating the Wnt signaling pathway. Cancer Cell Int 20:596 (2020). PubMed: 33317536
Bao C et al. Exploring specific prognostic biomarkers in triple-negative breast cancer. Cell Death Dis 10:807 (2019). PubMed: 31649243

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Key features and details

Related conjugates and formulations

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

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (8)

Customer reviews and Q&As