HNF-4-alpha Recombinant Rabbit Monoclonal Antibody [SN72-03]
cat.: ET1611-43
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: SN72-03
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 53 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human HNF-4-alpha aa 21-70 / 474.
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.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:2,000
1:100
1:200-1:3,000
1:1,000
Uniprot #: SwissProt: P41235 Human | P49698 Mouse | P22449 Rat
Alternative names: FLJ39654 FRTS4 Hepatic nuclear factor 4 alpha Hepatocyte nuclear factor 4 alpha Hepatocyte nuclear factor 4 Hepatocyte nuclear factor 4-alpha HNF 4 alpha HNF 4 HNF-4-alpha HNF4 HNF4A HNF4A_HUMAN HNF4a7 HNF4a8 HNF4a9 Hnf4alpha HNF4alpha10/11/12 MODY 1 MODY MODY1 NR2A1 NR2A21 Nuclear receptor subfamily 2 group A member 1 OTTHUMP00000031060 OTTHUMP00000031062 TCF 14 TCF TCF-14 TCF14 Tcf4 Transcription factor 14, hepatic nuclear factor Transcription factor 14 Transcription factor HNF 4 Transcription factor HNF-4 Transcription factor HNF4
Images
ET1611-43_1.jpg Fig1: 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.
ET1611-43_2.jpg Fig2: 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.
ET1611-43_3.jpg Fig3: 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.
ET1611-43_4.jpg Fig4: 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.
ET1611-43_5.jpg Fig5: 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) 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.
ET1611-43_6.jpg Fig6: 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) 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.
ET1611-43_7.jpg Fig7: 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) 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.
ET1611-43_8.jpg Fig8: 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) 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.
ET1611-43_9.jpg Fig9: 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).
ET1611-43_10.jpg Fig10: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.