Anti-FOXG1 抗体 [EPR18987]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunohistochemical analysis of paraffin-embedded rat E14 tissue labeling FOXG1 with ab196868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on cerebrum and olfactory epithelium of E14 rat is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunohistochemical analysis of formalin fixed paraffin embedded human glioma labelling FOXG1 with ab196868 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab196868 Anti-FOXG1 antibody [EPR18987] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling FOXG1 with ab196868 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in cells from a human glioma (PMID : 28465359) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-FOXG1 antibody [EPR18987] (AB196868)

FOXG1 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate with ab196868 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab196868 at 1/1000 dilution.

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

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-FOXG1 antibody [EPR18987] (ab196868) at 1/30 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab196868 in U-87 MG lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 55 kDa

true

Exposure time: 180s

• ChIP-seq

Lab

ChIP-sequencing - Anti-FOXG1 antibody [EPR18987] (AB196868)

Chromatin was prepared from U-87 MG cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab196868 [EPR18987]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-FOXG1 antibody [EPR18987] (AB196868)

Chromatin was prepared from U-87 MG cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab196868 [EPR18987]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-FOXG1 antibody [EPR18987] (AB196868)

Chromatin was prepared from U-87 MG cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab196868 [EPR18987]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilized frozen mouse embryo E14.5 tissue labeling FOXG1 with ab196868 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive nuclear staining in the cortical plate of the telencephalon (LV : lateral ventricle; PMID : 14704420) is observed. Counter stained with DAPI.

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 (green).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Intracellular flow cytometric analysis of4% paraformaldehyde fixed, 90% methanol permeabilized C6 (rat glial tumor cell line)cell line labeling FOXG1 with ab196868 at 1/60 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 Fluorr^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunohistochemical analysis of paraffin-embedded mouse E14 tissue labeling FOXG1 with ab196868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on cerebrum and olfactory epithelium of E14 mouse is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunofluorescent analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized C6 (rat glial tumor cell line) cells labeling FOXG1 with ab196868 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining on C6 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FOXG1 antibody [EPR18987] (AB196868)

Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilized frozen rat embryo E14.5 tissue labeling FOXG1 with ab196868 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive nuclear staining in the cortical plate of the telencephalon (LV : lateral ventricle; PMID : 14704420) is observed. Counter stained with DAPI.

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 (green).

• ChIP-seq

Lab

ChIP-sequencing - Anti-FOXG1 antibody [EPR18987] (AB196868)

Chromatin was prepared from Neuro-2a cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab196868 [EPR18987]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Unknown

Immunoprecipitation - Anti-FOXG1 antibody [EPR18987] (AB196868)

FOXG1 was immunoprecipitated from 0.35 mg mouse brain lysate 10 μg with ab196868 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab196868 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : mouse brain lysate 10 μg

Lane 2 : ab196868 IP in mouse brain cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab196868 in mouse brain lysate

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

Exposure time : 32s

All lanes:

Immunoprecipitation - Anti-FOXG1 antibody [EPR18987] (ab196868)

Predicted band size: 52 kDa

false

• ChIP-seq

Lab

ChIP-sequencing - Anti-FOXG1 antibody [EPR18987] (AB196868)

Chromatin was prepared from Neuro-2a cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab196868 [EPR18987]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-FOXG1 antibody [EPR18987] (AB196868)

Chromatin was prepared from Neuro-2a cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab196868 [EPR18987]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• WB

Supplier Data

Western blot - Anti-FOXG1 antibody [EPR18987] (AB196868)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lanes 1-4 : 15 seconds; Lanes 5-12 : 3 minutes.

This target is expressed during brain development (PMID : 26896590, PMID : 7959731). The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

All lanes:

Western blot - Anti-FOXG1 antibody [EPR18987] (ab196868) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Lane 4:

Human fetal spleen lysate at 10 µg

Lane 5:

Mouse brain lysate at 10 µg

Lane 6:

Mouse heart lysate at 10 µg

Lane 7:

Mouse kidney lysate at 10 µg

Lane 8:

Mouse spleen lysate at 10 µg

Lane 9:

Rat brain lysate at 10 µg

Lane 10:

Rat heart lysate at 10 µg

Lane 11:

Rat liver lysate at 10 µg

Lane 12:

Rat spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 52 kDa

Observed band size: 58 kDa

true

• WB

Supplier Data

Western blot - Anti-FOXG1 antibody [EPR18987] (AB196868)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular mass observed is consistent with the literature (PMID : 26508630). The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

All lanes:

Western blot - Anti-FOXG1 antibody [EPR18987] (ab196868) at 1/1000 dilution

Lane 1:

Rat E18 brain lysate at 20 µg

Lane 2:

Mouse E12.5 brain lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 58 kDa

true

Exposure time: 3s