Invitrogen

FOXJ1 Monoclonal Antibody (2A5), eBioscience™

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

40 Published Figures

67 References

View all (12) FOXJ1 antibodies

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Datasheet

Protocols

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FOXJ1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry of paraffin-embedded mouse lung tissue with Anti-Human/Mouse FOXJ1 Purified followed by Anti-Mouse IgG Biotin (Product # 13-4013-85) and Avidin HRP (Product # 18-4100-51) (right).Data provided courtesy of Steven L.Brody.

FOXJ1 Antibody in Western Blot (WB) — Immunoblotting of mouse tracheal epithelial cells at a 1:500 dilution of Anti-Human/Mouse FOXJ1 Purified.Bands were visualized using Anti-Mouse IgG HRP.

FOXJ1 Antibody — Lentiviral-mediated knock down of Syt-1 reduces endogenous A? levels in mouse primary neuronal culture. a Western blot analysis of APP CTF levels in mouse primary neurons infected with lentiviral shRNA against Syt-1. Reduction in the endogenous expression of Syt-1 was confirmed using a Syt-1 specific antibody (Synaptic Systems) while GAPDH staining was used to ensure equal protein loading. b Quantitative analysis of A?40 and A?42 levels from the conditioned media of mouse primary neurons infected with a control vector and lentiviral vector expressing Syt-1 shRNA. Lentiviral-mediated knock down of endogenous Syt-1 in mouse primary neuronal cultures significantly decreases secreted A?40 and A?42 levels as compared to the control vector (student t test; *, p < 0.05; n = 3 for each condition) {KD}

FOXJ1 Antibody in Immunocytochemistry (ICC/IF) — Figure 1 BMP signaling declines during ependymal cell differentiation. ( a ) Immunohistofluorescence analysis of Id1 in the LGE of the embryonic mouse brain at E14.5 or E16.5 as well as of Id1, glial fibrillary acidic protein (GFAP), and S100 in the SVZ of the lateral ventricles in the adult mouse brain (P56-P63). The boxed regions of the upper panels are shown at higher magnification in the corresponding lower panels. Arrowheads indicate Id1 + cells. Scale bars, 200 mum (upper panels) or 20 mum (lower panels). Data are representative of three independent experiments. ( b ) Quantification by immunohistofluorescence analysis of Id1 + cells among Sox2 + Ascl1 - NPCs in the LGE at E14.5 and E16.5. Data are means +- SEM (n = 3 embryos for each stage). Three sections were counted per sample. Total of 500-800 cells were counted per section. ** p < 0.01 (two-tailed Student''s unpaired t test). ( c ) Immunoblot analysis of the VZ-SVZ of the embryonic GE and corresponding postnatal basal ganglia at the indicated embryonic, postnatal, and young adult (P30) stages with antibodies to phosphorylated (p) Smad1/5/8, to Id1, to Id4, and to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, loading control). Data are representative of three independent experiments, with each experiment analyzed several times. ( d ) Dissociated cells of the lateral ventricular wall of P0 mice were cultured first for 5 days in vitro (DIV) in the presence of 10% FBS (proliferation medium) and then for up to 5 DIV

FOXJ1 Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — Figure 6. Expressions of Ano1 and Cftr in embryonic and fetal airways. ( A ) Expression of Shh , Cftr , and Ano1 in E15 trachea examined by FISH. Shh marks tracheal epithelial cells. Nuclei are stained by DAPI (white). Scale bar indicates 20 mu m. ( B ) Expression of Cftr and Ano1 in tracheal epithelial cells at different time points, with a cell type breakdown for E15 and E16. Expressions of Cftr and Ano1 in neonatal tracheal epithelial cells are shown in Figure 6--figure supplement 1 . ( C ) Expression of CFTR and ANO1 in human fetal trachea of GW 15 by FISH. Two areas (I: surface epithelium/SE; II: submucosal glands/SMG) of the fetal tracheal epithelium are enlarged. Nuclei are stained by DAPI (blue). Scale bar indicates 50 mu m. ( D ) Immunofluorescence staining of TP63 (green) and FOXJ1 (magenta) in human fetal tracheal sample at GW 15. SE and SMG are shown. Ciliated cells are only present in the surface epithelial cell layer. Epithelial cells are marked by E-cad in white. Nuclei are stained by DAPI (blue). Scale bar indicates 50 mu m. Figure 6--figure supplement 1. Expansion of the secretory cell lineage in global and conditional Ano1 knockouts. ( A ) Cellular composition for tracheal epithelial cells of wild-type littermates and Ano1 -/- mutants from different time points. ( B ) scGPS showing cell-cycle gene expression in every epithelial cell state at E16 in wild-type and Ano1 -/- mutant embryos. scGPS (single-cell Geneset Percentile Score) is an expression enrichment

FOXJ1 Antibody in Immunocytochemistry, Flow Cytometry (ICC/IF, Flow) — Figure 4 Loss of FGFR1 Signaling Results in a Block in Ciliated Cell Differentiation (A) Confocal image of Tg ( KRT5-CreER ); Rosa26R fGFP/+ ; Fgfr1 Delta/fx tracheal section. Green, GFP (reporter); blue, FGFR1; red, SCGB1A1 (secretory cells); white, acetylated tubulin (ACT, cilia). In the patch of GFP + cells marked by the dashed bracket, FGFR1 is absent and there are no ciliated cells. In the GFP + cells within the solid bracket, FGFR1 has not been deleted, there are fewer GFP + cells, and ciliated cell differentiation has occurred. (B) Percentage of GFP + cells that co-express SCGB1A1, ACT, or FOXJ1 in control Tg ( KRT5-CreER ); Rosa26R fGFP/+ ; Fgfr1 +/fx and cKO Tg ( KRT5-CreER ); Rosa26R fGFP/+ ; Fgfr1 Delta/fx animals 24 weeks after tmx administration. (C) Percentage of GFP + luminal cells in cKO Tg ( KRT5-CreER ); Rosa26R fGFP/+ ; Fgfr1 Delta/fx trachea that retain FGFR1 protein 24 weeks after tmx administration. (D) Sections from control and Fgfr1 cKO tracheae at 24 weeks after tmx administration. Green, GFP (reporter); red, gamma-tubulin (basal bodies); white, FOXJ1 (ciliated cells). (E) Schematic of in vitro experiment. (F) Day-11 ALI cultures grown from control and Fgfr1 cKO animals. Green, GFP (reporter); red, ACT (cilia); blue, DAPI (nuclei). Error bars denote SEM. Scale bars represent 100 mum in (A) and (F), and 50 mum in (D).

FOXJ1 Antibody in Immunocytochemistry, Immunohistochemistry (Frozen) (ICC/IF, IHC (F)) — Extended Data Figure 2 Ablation of ciliated cells has no effect on airway cell proliferation, mesenchymal cell types, mesenchymal morphology, and airway stem and progenitor cell replication over time a, Immunostaining for basal (CK5 (green)), ciliated (FOXJ1 (red)), and secretory cells (SCGB1A1 (white)) on either control (left panels) or tamoxifen (Tam)-treated FOXJ1-DTA mice (right panels) 5, 15, 30, 45, 60, and 150 days after ciliated cell ablation (n=3 mice). b, Quantification of absolute cell numbers of basal CK5 + cells (top graph) and secretory SCGB1A1 + cells (bottom graph) per trachea on control (black bars) or Tam-treated (white bars) mice over time (n=3 mice). c, Immunostaining for ciliated cells (FOXJ1 (green)) and proliferating cells (Ki67 (red)) on either control (upper panel) or tamoxifen (Tam)-treated FOXJ1-DTA mice (lower panel) (n=6 mice). On the right, quantification of the percentage of Ki67 + cells per total DAPI + cells in tracheal sections from control (C) or Tam-treated mice 3 days after cell ablation (n=3 mice). d, Quantification of the percentage of proliferating Ki67 + cells relative to total DAPI + cells in control (black bars) or Tam-treated (white bars) mice over time (n=3 mice). e, Immunostaining for ciliated cells (ACTUB (green)) and cells that have undergone proliferation (BrdU (red)) on either control (upper panel) or Tam-treated mice (lower panel) at day 3 (n=6 mice). On the right, quantification of the percentage of BrdU + cells per total DA

FOXJ1 Antibody in Immunocytochemistry, Immunohistochemistry, Immunohistochemistry (PFA fixed) (ICC/IF, IHC, IHC (PFA)) — Figure 5 Dedifferentiated cells are functional stem cells both in vivo and ex vivo a, Schematic representation of the regeneration of epithelium from dedifferentiated cells after SO2 or influenza induced injury. b, Timeline for the induction of dedifferentiation prior to infectious or toxic injury followed by tissue harvest. c, Co-labeling of YFP (green) with CK5 (red; left panels), SCGB1A1 (red; middle panels) and FoxJ1 (red; right panels) after SO 2 (upper panels) or influenza induced injury (lower panels). d. Ex vivo expansion, and differentiation of sorted dedifferentiated cells. e, Immunostaining for CK5(red), p63 (magenta) and YFP (green) on colonies from sorted dedifferentiated cells. f, Co-labeling of YFP (green) with p63 or Acetylated-Tubulin or SCGB1A1 (red). i-Dox, inhaled doxycycline; Tam, tamoxifen; H, harvest. Nuclei - DAPI (blue). n=3 (2 replicates/mice per condition). Scale bar 20um.

FOXJ1 Antibody in Immunohistochemistry (IHC) — FIGURE 1 ID4 expression in ependymal cells. (A) Scheme of a coronal section of a mouse brain hemisphere showing the ventricular-subventricular zone (V-SVZ) of the lateral ventricles (LV) and a wholemount section that can be dissected out from this region. (B) Immunolabeling of ID4 and FOXJ1 (ependymal cell marker) on wholemount sections of adult C57BL/6J mice. Scale bar: 5 mum. (C) Immunogold staining of ID4 protein on V-SVZ sections. Left panel shows ependymal cells next to neural progenitors and neuroblasts (identified by pseudo-colors). Arrowheads indicate some of the gold particles labeling ID4 protein in ependymal cells. Right panel shows higher magnification of boxed area on left panel. Scale bar: 5 mum left and 2 mum right.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1-10 µg/mL

View 3 publications 3 publications

Immunohistochemistry (IHC)

View 35 publications 35 publications

Immunohistochemistry (Paraffin) (IHC (P))

1-10 µg/mL

View 2 publications 2 publications

Immunohistochemistry (PFA fixed) (IHC (PFA))

View 1 publication 1 publication

Immunohistochemistry (Frozen) (IHC (F))

View 3 publications 3 publications

Immunocytochemistry (ICC/IF)

Assay-Dependent

View 20 publications 20 publications

Flow Cytometry (Flow)

View 2 publications 2 publications

ChIP assay (ChIP)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Mouse

Published species

Fruit fly, Human, Mouse, Pig, Rat

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

2A5

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_1548835

Product Specific Information

Description: This 2A5 monoclonal antibody reacts with Foxj1, a 50-kDa member of the Forkhead-box (FOX) family of winged-helix transcription factors. Expression of Foxj1 is high in ciliated epithelial cells and low in naive T and B cells, where cellular activation results in downregulation of the factor. Foxj1 modulates germinal center formation and Th1 activation by repressing NFkappaB through IkappaBbeta induction. As such, this transcription factor has been suggested to antagonize autoimmune reactions such as systemic lupus erythematosus and rheumatoid arthritis. In addition, Foxj1 has been linked to the formation of motile cilia in epithelial cells.

Applications Reported: This 2A5 antibody has been reported for use in western blotting, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, and immunocytochemistry.

Applications Tested: This 2A5 antibody has been tested by western blot of mouse tracheal epithelial cells and testes, as well as by immunohistochemistry of mouse lung tissue. This antibody can be used between 1-10 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Forkhead-box J1 (FOXJ1) is a 421-amino acid transcription factor that suppresses T cell activity and spontaneous autoimmunity, through the repression of NF B activity. FOXJ1 also inhibits the humoral immune response in B cells. FOXJ1 deficiency in B cells results in spontaneous and accentuated germinal center formation and is implicated in the development of pathogenic autoantibodies and accentuated responses to immunizations. Abnormal expression of FOXJ1 may be associated with autoimmune diseases and/or other inflammatory diseases. FOXJ1 is also required for cilia formation and left-right axis determination because it increases calpastatin expression, a protein necessary for the ability of basal bodies to anchor to the apical cytoskeleton. FOXJ1 expression may function as an early marker of epithelial cell differentiation, recovery, and function.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: fork head homologue 4; Forkhead box protein J1; forkhead transcription factor HFH-4; forkhead-like 13; Forkhead-related protein FKHL13; Hepatocyte nuclear factor 3 forkhead homolog 4; HFH-4; HNF-3/forkhead homolog 4; MGC35202

Gene Aliases: FKHL-13; FKHL13; FOXJ1; HFH-4; HFH4

UniProt ID: (Human) Q92949, (Mouse) Q61660

Entrez Gene ID: (Human) 2302, (Mouse) 15223

Function(s)