Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16885-99] to HNF-4-alpha
Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-HNF-4-alpha antibody [EPR16885-99]
See all HNF-4-alpha primary antibodies
Description

Rabbit monoclonal [EPR16885-99] to HNF-4-alpha
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Mouse Rat Human
WB	Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HepG2 and Caco-2 whole cell lysates; Human colon and fetal kidney lysates. IHC-P: Human colon, Human liver, mouse liver and rat colon tissues. ICC/IF: HepG2 and HT-29 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16885-99
Isotype

IgG
Research areas

Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution + Full length mouse HNF-4-gamma recombinant protein at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 53 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Based on sequence analysis of the immunogen, ab201460 shares 72% homology with HNF4 gamma. The levels of XR were tested in the accompanying recombinant protein, WB experiment.
Flow Cytometry - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

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.
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

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.
Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/10000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution + Caco-2 (Human colorectal adenocarcinoma cells) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

All lanes : Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution

Lane 1 : Human colon lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

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