Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution (green).

Confocal image showing nuclear staining on HT-29 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is stained with ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
-ve control 1 : ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
-ve control 2 : ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201460).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) labelling CDKN2A/p16INK4a with purified ab201460 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Alexa Fluor^® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201460).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

This IHC data was generated using the same anti-HNF4 antibody clone, EPR16885-99, in a different buffer formulation (cat# ab201460).

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on Human colon tissue 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) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on Human liver tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201460).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HT-29 (Human colorectal adenocarcinoma cells) cells labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution (green).

Confocal image showing nuclear staining on HT-29 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is stained with ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
-ve control 1 : ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
-ve control 2 : ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201460).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

This IHC data was generated using the same anti-HNF4 antibody clone, EPR16885-99, in a different buffer formulation (cat# ab201460).

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on rat colon tissue 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) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Nuclear staining on mouse liver tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201460).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (AB231167)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of ab201460 [EPR16885-99)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201460).