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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade - BSA and Azide free (AB219610)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of Human colon 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 (ab181604).

Perform heat mediated antigen retrieval 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] - ChIP Grade - BSA and Azide free (AB219610)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human liver 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 (ab181604).

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

• ChIP

Supplier Data

ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade - BSA and Azide free (AB219610)

Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab181604 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

ABCC6_NC1 is negative control

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181604).

• IP

Supplier Data

Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade - BSA and Azide free (AB219610)

HNF-4 alpha was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell extract with ab181604 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab181604 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HepG2 whole cell extract 10 µg (Input).

Lane 2 : ab181604 IP in HepG2 whole cell extract.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab181604 in HepG2 whole cell extract.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181604).

All lanes:

Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hnf-4-alpha-antibody-epr16885-chip-grade-ab181604'>ab181604</a>)

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

Exposure time: 5s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade - BSA and Azide free (AB219610)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab181604 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 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 (ab181604).

Perform heat mediated antigen retrieval 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] - ChIP Grade - BSA and Azide free (AB219610)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of rat colon 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 (ab181604).

Perform heat mediated antigen retrieval 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] - ChIP Grade - BSA and Azide free (AB219610)

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 ab181604 [EPR16885]. 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 downloaded here. 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 (ab181604).