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HNF-4-alpha Recombinant Rabbit Monoclonal Antibody [SN72-03]

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Specification

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Catalog# ET1611-43

HNF-4-alpha Recombinant Rabbit Monoclonal Antibody [SN72-03]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

HNF-4-alpha Recombinant Rabbit Monoclonal Antibody [SN72-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human HNF-4-alpha aa 21-70 / 474.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 53 kDa

Positive Control

A549 cell lysates, LOVO cell lysates, HepG2, SW480, human colon cancer tissue, human small intestine tissue, mouse colon tissue, rat colon tissue, Mouse liver tissue lysate, Mouse kidney tissue lysate, Mouse brain tissue lysate.

Conjugation

unconjugated

Clone Number

SN72-03

RRID

AB_3070022

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200-1:3,000

  • FC

  • 1:1,000

Target

Function

HNF-4 alpha is a transcription factor that binds DNA as a homodimer. HNF-4 alpha is important in liver, kidney, and intestinal development. It has also been intensely studied as one of a variety of genes responsible for diabetes mellitus. HNF-4 alpha has been shown in knock out mice to be essential for the morphogenic and functional differentiation of hepatocytes. HNF-4 alpha is a dominant regulator of epithelial phenotypes able to drive the mesenchymal-to-epithelial transition when expressed in fibroblasts.

Background References

1. Simeone P et al. A unique four-hub protein cluster associates to glioblastoma progression. PLoS One 9:e103030 (2014).

2. Saha SK et al. Mutant IDH inhibits HNF-4a to block hepatocyte differentiation and promote biliary cancer. Nature 513:110-4 (2014).

Sequence Similarity

Belongs to the nuclear hormone receptor family. NR2 subfamily.

Post-translational Modification

Phosphorylated on tyrosine residue(s); phosphorylation is important for its DNA-binding activity. Phosphorylation may directly or indirectly play a regulatory role in the subnuclear distribution. Phosphorylation at Ser-313 by AMPK reduces the ability to form homodimers and bind DNA.; Acetylation at Lys-458 lowers transcriptional activation by about two-fold.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P41235 Human

SwissProt: P49698 Mouse

SwissProt: P22449 Rat

Synonyms

FLJ39654 antibody

FRTS4 antibody

Hepatic nuclear factor 4 alpha antibody

Hepatocyte nuclear factor 4 alpha antibody

Hepatocyte nuclear factor 4 antibody

Hepatocyte nuclear factor 4-alpha antibody

HNF 4 alpha antibody

HNF 4 antibody

HNF-4-alpha antibody

HNF4 antibody

Expand

Images

  • Western blot analysis of HNF-4-alpha on A549 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of HNF-4-alpha on A549 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-43, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of HNF-4-alpha on LOVO cell lysates with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/500 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 53 kDa<br />Observed band size: 53 kDa<br /><br />Exposure time: 2 minutes;<br /><br />10% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of HNF-4-alpha on LOVO cell lysates with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/500 dilution.

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 53 kDa
    Observed band size: 53 kDa

    Exposure time: 2 minutes;

    10% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-43) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling HNF-4-alpha with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling HNF-4-alpha with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of SW480 cells labeling HNF-4-alpha with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SW480 cells labeling HNF-4-alpha with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/3,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/3,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-43) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-43) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/3,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/3,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-43) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/3,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/3,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-43) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HepG2 cells labeling HNF-4-alpha.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling HNF-4-alpha.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-43, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Western blot analysis of HNF-4-alpha on different lysates with Rabbit anti-HNF-4-alpha antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse liver tissue lysate<br />Lane 2: Mouse kidney tissue lysate<br />Lane 3: Mouse brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 53 kDa<br />Observed band size: 53 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-43" style="font-weight: bold;text-decoration: underline;">ET1611-43</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of HNF-4-alpha on different lysates with Rabbit anti-HNF-4-alpha antibody (ET1611-43) at 1/1,000 dilution.

    Lane 1: Mouse liver tissue lysate
    Lane 2: Mouse kidney tissue lysate
    Lane 3: Mouse brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 53 kDa
    Observed band size: 53 kDa

    Exposure time: 3 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-43) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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