Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)

Rabbit Recombinant Monoclonal hnRNP D/AUF1 antibody. Suitable for ICC/IF, IP, Flow Cyt (Intra), WB, IHC-P and reacts with Mouse, Human, Rat samples. Cited in 2 publications.

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Images

Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (AB259895), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	Flow Cyt (Intra)	WB	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Tested	Tested	Tested
Rat	Expected	Expected	Expected	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information HNRNPD

Function

Binds with high affinity to RNA molecules that contain AU-rich elements (AREs) found within the 3'-UTR of many proto-oncogenes and cytokine mRNAs. Also binds to double- and single-stranded DNA sequences in a specific manner and functions a transcription factor. Each of the RNA-binding domains specifically can bind solely to a single-stranded non-monotonous 5'-UUAG-3' sequence and also weaker to the single-stranded 5'-TTAGGG-3' telomeric DNA repeat. Binds RNA oligonucleotides with 5'-UUAGGG-3' repeats more tightly than the telomeric single-stranded DNA 5'-TTAGGG-3' repeats. Binding of RRM1 to DNA inhibits the formation of DNA quadruplex structure which may play a role in telomere elongation. May be involved in translationally coupled mRNA turnover. Implicated with other RNA-binding proteins in the cytoplasmic deadenylation/translational and decay interplay of the FOS mRNA mediated by the major coding-region determinant of instability (mCRD) domain. May play a role in the regulation of the rhythmic expression of circadian clock core genes. Directly binds to the 3'UTR of CRY1 mRNA and induces CRY1 rhythmic translation. May also be involved in the regulation of PER2 translation.

Alternative names

AUF1, HNRPD, HNRNPD, Heterogeneous nuclear ribonucleoprotein D0, hnRNP D0, AU-rich element RNA-binding protein 1

Recommended products

Rabbit Recombinant Monoclonal hnRNP D/AUF1 antibody. Suitable for ICC/IF, IP, Flow Cyt (Intra), WB, IHC-P and reacts with Mouse, Human, Rat samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR24001-12
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The hnRNP D/AUF1 protein also known as heterogeneous nuclear ribonucleoprotein D has a molecular mass of approximately 37-40 kDa. It plays a significant role in the cellular machinery by binding to RNA molecules mainly impacting mRNA stability and turnover. The protein is expressed ubiquitously in various tissues throughout the body indicating its importance in numerous cellular functions. It is one of the hnRNP family members involved in the post-transcriptional regulation of gene expression.

Biological function summary

HnRNP D/AUF1 regulates the degradation of mRNA by binding to AU-rich elements (AREs) found in the 3' untranslated region of many genes. It is an integral part of RNA-protein complexes that are important for controlling the half-life of mRNAs. Besides mRNA decay hnRNP D/AUF1 participates in other processes like mRNA splicing and transport. Its interactions within the ribonucleoprotein complexes highlight its versatile role in RNA metabolism.

Pathways

HnRNP D/AUF1 operates within important biological processes such as the mRNA decay pathway and the stress response pathway. It interacts closely with other proteins involved in mRNA decay such as tristetraprolin and members of the poly-A binding protein family. hnRNP D/AUF1 influences the decay rate of different transcripts contributing to cellular responses to environmental stimuli and maintaining homeostasis.

Associated diseases and disorders

HnRNP D/AUF1 links to inflammatory conditions and some types of cancer. Altered expression levels of AUF1 have been associated with chronic inflammation where its role in mRNA stability impacts the expression of cytokines. In cancer dysregulation of AUF1 can lead to incorrect mRNA turnover of tumor suppressors or oncogenes linking it to proteins like p53 in the context of tumorigenesis. These associations make hnRNP D/AUF1 a potential target for therapeutic interventions in related diseases.

Product promise

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Full details and terms and conditions can be found here:
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12 product images

Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
False colour image of Western blot: Anti-hnRNP D/AUF1 antibody [EPR24001-12] staining at 1/1000 dilution, shown in green; loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab259895 was shown to bind specifically to hnRNP D/AUF1. A band was observed at 40 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in HNRNPD knockout cell line Human HNRNPD knockout U-2 OS cell line ab273860 (knockout cell lysate Human HNRNPD knockout U-2 OS cell lysate ab273814). To generate this image, wild-type and HNRNPD knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895) at 1/1000 dilution
Lane 1: Wild-type U-2 OS cell lysate at 20 µg
Lane 2: HNRNPD knockout U-2 OS cell lysate at 20 µg
Lane 2: Western blot - Human HNRNPD knockout U-2 OS cell line (Human HNRNPD knockout U-2 OS cell line ab273860)
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 40 kDa
Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes
All lanes: Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 3: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
Lane 5: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 38 kDa
Observed band size: 37 kDa, 42 kDa
Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 81 seconds
All lanes: Western blot - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 40 µg
Lane 2: C6 (rat glial tumor glial cell) whole cell lysate at 40 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 38 kDa
Observed band size: 42 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling hnRNP D/AUF1 with ab259895 at 1/100 (5.59 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human breast. The section was incubated with ab259895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labelling hnRNP D/AUF1 with ab259895 at 1/100 (5.59 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human lung carcinoma. The section was incubated with ab259895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling hnRNP D/AUF1 with ab259895 at 1/100 (5.59 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse cerebrum. The section was incubated with ab259895 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling hnRNP D/AUF1 with ab259895 at 1/50 (11.18 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing nuclear staining in HeLa cells is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Immunoprecipitation - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
hnRNP D/AUF1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug with ab259895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug
Lane 2: ab259895 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259895 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
All lanes: Immunoprecipitation - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Predicted band size: 38 kDa
Observed band size: 42 kDa
Immunocytochemistry/ Immunofluorescence - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling hnRNP D/AUF1 with ab259895 at 1/50 (11.18 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing nuclear staining in NIH/3T3 cells is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Immunoprecipitation - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
hnRNP D/AUF1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab259895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: ab259895 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259895 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
All lanes: Immunoprecipitation - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Predicted band size: 38 kDa
Observed band size: 42 kDa
Flow Cytometry (Intracellular) - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma cell) cells labelling hnRNP D/AUF1 with ab259895 at 1/500 dilution (0.1ug)/ (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-hnRNP D/AUF1 antibody [EPR24001-12] (ab259895)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling hnRNP D/AUF1 with ab259895 at 1/500 dilution (0.1ug)/(Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.