Mouse Anti-ALYREF Recombinant Antibody (11G5) (CBMAB-T0037-YJ)

Name: Mouse Anti-ALYREF Recombinant Antibody (11G5)
SKU: CBMAB-T0037-YJ
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Published Data

Mouse Anti-ALYREF Recombinant Antibody (11G5) in IF

Colocalization of PRRSV nsp1β. HeLa cells were transfected with FLAG-nsp1β plasmid and stained with anti-ALY MAb (green). ...View More

Ke, H., Han, M., Kim, J., Gustin, K. E., & Yoo, D. (2019). Porcine reproductive and respiratory syndrome virus nonstructural protein 1 beta interacts with nucleoporin 62 to promote viral replication and immune evasion. Journal of virology, 93(14), 10-1128.

Mouse Anti-ALYREF Recombinant Antibody (11G5) in IF

Four-cell stage siControl, siIws1, and siSupt6 embryos were double-stained for Alyref and Pol II CTD Ser2p. PLA shows that either Iws1 or Supr6 is required for the interaction of Alyref with Pol II CTD Ser2p. Scale bars: 20 μm. ...View More

Oqani, R. K., Lin, T., Lee, J. E., Kang, J. W., Shin, H. Y., & Il Jin, D. (2019). Iws1 and Spt6 regulate trimethylation of histone H3 on lysine 36 through Akt signaling and are essential for mouse embryonic genome activation. Scientific reports, 9(1), 3831.

Mouse Anti-ALYREF Recombinant Antibody (11G5) in IP

ZC3H14 interacts with the THO complex. (A) Co-immunoprecipitation of ZC3H14 and THO components from mouse brain lysate. The Input and immunoprecipitated samples for ZC3H14, ZC3H14 + RNase, and control IgG are shown. THOC1 and THOC2 serve as representative members of the THO complex, together with the THO-associated protein ALYREF. HuR serves as a negative control. Asterisks indicate non-specific protein bands detected by the commercial antibodies. (B) Reciprocal co-immunoprecipitation using anti-THOC1. The Input and immunoprecipitated samples for THOC1, THOC1 + RNase and control IgG immunoprecipitations were probed to detect THOC1, ZC3H14 and THOC2 proteins. (C) Sedimentation of ZC3H14 protein complexes from the nuclear fraction of mouse brain lysate through a 10-50% glycerol gradient. Gradient fractions from top to bottom were resolved on SDS-PAGE from left to right. Fractions were probed with antibodies to detect ZC3H14, THOC1, THOC2, ALYREF and Nucleophosmin (NPM1) as a control. The Input sample is shown on the left. Overlapping fractions are outlined. Molecular weights are shown in kDa and indicated to the left. ...View More

Morris, K. J., & Corbett, A. H. (2018). The polyadenosine RNA-binding protein ZC3H14 interacts with the THO complex and coordinately regulates the processing of neuronal transcripts. Nucleic acids research, 46(13), 6561-6575.

Mouse Anti-ALYREF Recombinant Antibody (11G5) in WB

Western blot of NORAD RAP experiments with or without DNA damage. Western blots are representative of one experiment; three independent experiments were performed. ...View More

Munschauer, M., Nguyen, C. T., Sirokman, K., Hartigan, C. R., Hogstrom, L., Engreitz, J. M., ... & Lander, E. S. (2018). The NORAD lncRNA assembles a topoisomerase complex critical for genome stability. Nature, 561(7721), 132-136.

Datasheet

SDS

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Datasheet Target References Q & As Review & reward Protocols Associated Products

Basic Information

Host Animal

Mouse

Clone

11G5

Application

IHC-P, IP, WB, IF

Immunogen

Recombinant human ALY/REF fusion protein.

Host Species

Mouse

Specificity

Human, Mouse, Rat, Zebrafish

Antibody Isotype

IgG1, κ

Clonality

Monoclonal

Application Notes

The COA includes recommended starting dilutions, optimal dilutions should be determined by the end user.

Application	Note
WB	1:100-1:1,000
IP	1-2 µg per 100-500 µg of total protein (1 ml of cell lysate)
IF(ICC)	1:50-1:500
IHC	1:50-1:500

Formulations & Storage [For reference only, actual COA shall prevail!]

Format

Liquid

Buffer

PBS, 0.1% gelatin

Preservative

< 0.1% sodium azide

Concentration

0.2 mg/ml

Storage

Store at 4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

More Infomation

Target

Full Name

Aly/REF Export Factor

Introduction

ALYREF is a heat stable, nuclear protein and plays a role as a molecular chaperone. ALYREF is thought to regulate dimerization, DNA binding, and transcriptional activity of basic region-leucine zipper (bZIP) proteins.

Entrez Gene ID

Human	10189
Frog	549781

UniProt ID

Human	Q86V81
Frog	Q28FB9

Alternative Names

Aly/REF Export Factor; Transcriptional Coactivator Aly/REF; BZIP-Enhancing Factor BEF; Ally Of AML-1 And LEF-1; THO Complex 4; THOC4; Tho4

Function

Export adapter involved in nuclear export of spliced and unspliced mRNA. Binds mRNA which is thought to be transferred to the NXF1-NXT1 heterodimer for export (TAP/NFX1 pathway) (PubMed:15833825, PubMed:15998806, PubMed:17190602, PubMed:11707413, PubMed:11675789, PubMed:11979277, PubMed:18364396, PubMed:22144908, PubMed:22893130, PubMed:23222130, PubMed:25662211). Component of the 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 (PubMed:15833825, PubMed:15998806, PubMed:17190602). TREX recruitment occurs via an interaction between ALYREF/THOC4 and the cap-binding protein NCBP1 (PubMed:15833825, PubMed:15998806, PubMed:17190602). 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 (PubMed:18974867). Required for TREX complex assembly and for linking DDX39B to the cap-binding complex (CBC) (PubMed:15998806, PubMed:17984224). 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 the nuclear export of intronless mRNA; proposed to be recruited to intronless mRNA by ATP-bound DDX39B. Involved in transcription elongation and genome stability (PubMed:12438613, PubMed:17984224). 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 chaperone and promotes the dimerization of transcription factors containing basic leucine zipper (bZIP) domains and thereby promotes transcriptional activation.

Biological Process

mRNA 3'-end processing Source: Reactome
mRNA export from nucleus Source: UniProtKB
mRNA splicing, via spliceosome Source: Reactome
Osteoblast differentiation Source: UniProtKB
Positive regulation of DNA-templated transcription, elongation Source: UniProtKB
Regulation of DNA recombination Source: UniProtKB
Replication fork processing Source: UniProtKB
RNA export from nucleus Source: UniProtKB
Viral mRNA export from host cell nucleus Source: UniProtKB
Viral process Source: UniProtKB-KW

Cellular Location

Nucleus; Nucleus speckle; Cytoplasm. Colocalizes with the core EJC, ALYREF/THOC4, NXF1 and DDX39B in the nucleus and nuclear speckles. Travels to the cytoplasm as part of the exon junction complex (EJC) bound to mRNA (PubMed:19324961). Localizes to regions surrounding nuclear speckles known as perispeckles in which TREX complex assembly seems to occur (PubMed:23826332).

PTM

Arg-204 is dimethylated, probably to asymmetric dimethylarginine. Arginine methylation reduces RNA binding.2 Publications
Citrullinated by PADI4.

Wang, J. Z., Zhu, W., Han, J., Yang, X., Zhou, R., Lu, H. C., ... & Yang, H. (2021). The role of the HIF‐1α/ALYREF/PKM2 axis in glycolysis and tumorigenesis of bladder cancer. Cancer Communications.

Wang, J., Li, Y., Xu, B., Dong, J., Zhao, H., Zhao, D., & Wu, Y. (2021). ALYREF Drives Cancer Cell Proliferation Through an ALYREF-MYC Positive Feedback Loop in Glioblastoma. OncoTargets and therapy, 14, 145.

Qiao, L., Xu, L., Yu, L., Wynn, J., Hernan, R., Zhou, X., ... & Chung, W. K. (2021). Rare and de novo variants in 827 congenital diaphragmatic hernia probands implicate LONP1 and ALYREF as new candidate risk genes. medRxiv.

Tunnicliffe, R. B., Tian, X., Storer, J., Sandri-Goldin, R. M., & Golovanov, A. P. (2018). Overlapping motifs on the herpes viral proteins ICP27 and ORF57 mediate interactions with the mRNA export adaptors ALYREF and UIF. Scientific reports, 8(1), 1-12.

Yang, X., Yang, Y., Sun, B. F., Chen, Y. S., Xu, J. W., Lai, W. Y., ... & Yang, Y. G. (2017). 5-methylcytosine promotes mRNA export—NSUN2 as the methyltransferase and ALYREF as an m 5 C reader. Cell research, 27(5), 606-625.

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Please try the standard protocols which include: protocols, troubleshooting and guide.

Enzyme-linked Immunosorbent Assay (ELISA)
Flow Cytometry
Immunofluorescence (IF)
Immunohistochemistry (IHC)
Immunoprecipitation (IP)
Western Blot (WB)
Enzyme Linked Immunospot (ELISpot)
Proteogenomic
Other Protocols

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For research use only. Not intended for any clinical use.

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