Anti-Aly/Ref antibody [EPR17942] (ab202894)

Anti-Aly/Ref antibody [EPR17942] (ab202894) is a rabbit monoclonal antibody that is used to detect Aly/Ref in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

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ab240364
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Anti-Aly/Ref antibody [EPR17942] - BSA and Azide free
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PE Anti-Aly/Ref antibody [EPR17942]
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APC Anti-Aly/Ref antibody [EPR17942]
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Alexa Fluor® 594 Anti-Aly antibody [EPR17942]
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Alexa Fluor® 568 Anti-Aly antibody [EPR17942]
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Alexa Fluor® 555 Anti-Aly antibody [EPR17942]
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HRP Anti-Aly/Ref antibody [EPR17942]
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Alexa Fluor® 488 Anti-Aly antibody [EPR17942]
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Alexa Fluor® 750 Anti-Aly/Ref antibody [EPR17942]

Images

Western blot - Anti-Aly/Ref antibody [EPR17942] (AB202894), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% 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

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Expected	Expected	Tested
Rat	Expected	Tested	Tested	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/40	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 Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/2000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/250	Notes -
Species Human	Dilution info 1/250	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/150	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information ALYREF

Function

Functions as an mRNA export adapter; component of the transcription/export (TREX) complex which is thought to couple mRNA transcription, processing and nuclear export, and specifically associates with spliced mRNA and not with unspliced pre-mRNA (PubMed:15833825, PubMed:15998806, PubMed:17190602). TREX is recruited to spliced mRNAs by a transcription-independent mechanism, binds to mRNA upstream of the exon-junction complex (EJC) and is recruited in a splicing- and cap-dependent manner to a region near the 5' end of the mRNA where it functions in mRNA export to the cytoplasm via the TAP/NXF1 pathway (PubMed:15833825, PubMed:15998806, PubMed:17190602). Involved in the nuclear export of intronless mRNA; proposed to be recruited to intronless mRNA by ATP-bound DDX39B (PubMed:17984224). Plays a key role in mRNP recognition and mRNA packaging by bridging the mRNP-bound EJC and the TREX core complex (PubMed:37020021). TREX recruitment occurs via an interaction between ALYREF/THOC4 and the cap-binding protein NCBP1 (PubMed:15833825, PubMed:15998806, PubMed:17190602, PubMed:37020021). Required for TREX complex assembly and for linking DDX39B to the cap-binding complex (CBC) (PubMed:15998806, PubMed:17984224, PubMed:37020021). Binds mRNA which is thought to be transferred to the NXF1-NXT1 heterodimer for export (TAP/NXF1 pathway) (PubMed:11675789, PubMed:11707413, PubMed:11979277, PubMed:15833825, PubMed:15998806, PubMed:17190602, PubMed:18364396, PubMed:22144908, PubMed:22893130, PubMed:23222130, PubMed:25662211). In conjunction with THOC5 functions in NXF1-NXT1 mediated nuclear export of HSP70 mRNA; both proteins enhance the RNA binding activity of NXF1 and are required for NXF1 localization to the nuclear rim (PubMed:19165146). Involved in mRNA export of C5-methylcytosine (m5C)-containing mRNAs: specifically recognizes and binds m5C mRNAs and mediates their nucleo-cytoplasmic shuttling (PubMed:28418038). Acts as a chaperone and promotes the dimerization of transcription factors containing basic leucine zipper (bZIP) domains and thereby promotes transcriptional activation (PubMed:10488337). Involved in transcription elongation and genome stability (PubMed:12438613). (Microbial infection) The TREX complex is essential for the export of Kaposi's sarcoma-associated herpesvirus (KSHV) intronless mRNAs and infectious virus production; ALYREF/THOC4 mediates the recruitment of the TREX complex to the intronless viral mRNA.

Alternative names

ALY, BEF, THOC4, ALYREF, THO complex subunit 4, Tho4, Ally of AML-1 and LEF-1, Aly/REF export factor, Transcriptional coactivator Aly/REF, bZIP-enhancing factor BEF

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% 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: EPR17942
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

What is this antibody validated in?

Anti-Aly/Ref antibody [EPR17942] (ab202894) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.

What is the molecular weight of Aly/Ref?

Anti-Aly/Ref [EPR17942] (ab202894) specifically detects a band for Aly/Ref (UniProt: Q86V81) at a molecular weight of 27kDa.

Trusted by the scientific community

Anti-Aly/Ref [EPR17942] (ab202894) was first used in a scientific publication in 2015 and has been cited over 10 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR17942] also available for your convenience:
ab202894, Carrier free - ab240364, PE - ab305874, APC - ab305875, HRP - ab305876, Alkaline Phosphatase - ab308738, Alexa Fluor® 488 - ab309694, Alexa Fluor® 647 - ab310060, Alexa Fluor® 594 - ab310452, Alexa Fluor® 555 - ab311981, Alexa Fluor® 568 - ab312456, Alexa Fluor® 750 - ab321354

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

What are the advantages of a recombinant monoclonal antibody?
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

Aly/Ref also known as THOC4 is a protein that plays an important role in mRNA processing and transport. It is a member of the TREX (transcription/export) complex having a molecular mass of approximately 27 kDa. Aly/Ref is primarily expressed in the nucleus where it binds to spliced mRNAs and ensures their export to the cytoplasm. Functionally Aly/Ref interacts with mRNA export adapters attaching to the cap-binding complex at the 5' end of mRNA facilitating its journey to the nuclear pore.

Biological function summary

Aly/Ref functions by forming tight associations with the TREX complex which includes other proteins like UAP56. It is responsible for coupling transcription with mRNA processing and export helping to maintain the integrity of gene expression. Aly/Ref directly influences the efficiency and accuracy of mRNA export impacting gene regulation by ensuring that only fully processed transcripts exit the nucleus. By doing so it helps coordinate the transcription and mRNA export processes.

Pathways

Aly/Ref associates with the mRNA surveillance pathway and the RNA export pathway. Aly/Ref works closely with proteins like EJC components which are involved in checking mRNA quality before export. This participation ensures that only correctly spliced mRNAs move from the nucleus to the cytoplasm where they translate into proteins. Aly/Ref also contributes to maintaining cellular homeostasis by regulating the amount of mRNA available for translation.

Associated diseases and disorders

Aly/Ref relates to cancer and neurological disorders. Changes in the expression or function of Aly/Ref can disrupt mRNA export leading to abnormal protein synthesis which is associated with cancer development. Furthermore Aly/Ref interacts with proteins such as NXF1 which are implicated in the onset of certain neurological disorders when mutated or misregulated revealing its broader impact on cellular health and disease progression.

Product promise

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10 product images

Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/10000 dilution
Lane 1: PC-3 (Human prostate cancer cell line) whole cell lysate at 20 µg
Lane 2: HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg
Lane 3: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: HEK293 (Human epithelial cells from embryonic kidney) 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 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 5s
Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/2000 dilution
Lane 1: Human fetal brain lysate at 10 µg
Lane 2: Human fetal heart lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
Secondary
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15s
Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/2000 dilution
Lane 1: Mouse brain lysate at 10 µg
Lane 2: Mouse heart lysate at 10 µg
Lane 3: Rat brain lysate at 10 µg
Lane 4: Rat heart lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human cervix carcinoma 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) (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-Aly/Ref antibody [EPR17942] (ab202894)
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on mouse cerebral cortex 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) (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-Aly/Ref antibody [EPR17942] (ab202894)
Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on rat 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) (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.
Immunoprecipitation - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Aly/Ref was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab202894 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab202894 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 lysate 10 μg (Input). Lane 2: ab202894 IP in HepG2 whole cell lysate. Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab202894 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
All lanes: Immunoprecipitation - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Predicted band size: 27 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Aly/Ref with ab202894 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab202894 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cells) cells labeling Aly/Ref with ab202894 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on C6 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab202894 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Aly/Ref antibody [EPR17942] (ab202894)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Aly/Ref with ab202894 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.