XRN2 Antibody (Rabbit mAb) [J9K11]

Application: Reactivity: Human, Mouse, Rat

Usage Information

Dilution
WB1:1000 IHC1:1000 IF1:100 FCM1:500

Application
WB, IHC, IF, FCM

Reactivity
Human, Mouse, Rat

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
109 kDa 108 kDa,36 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Mouse lung tissue; Human lung tissue; Human cerebrum tissue; Rat cerebrum tissue; Mouse cerebrum tissue; Human cerebellum tissue; Mouse skeletal muscle tissue; Mouse brain tissue; Rat skeletal muscle tissue; Rat brain tissue; Human liver tissue; HeLa cells; NIH/3T3 cells; LN-229 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
XRN2 Antibody (Rabbit mAb) [J9K11] detects endogenous levels of total XRN2 protein.

Clone
J9K11

Synonym(s)
5'-3' exoribonuclease 2, DHM1-like protein, DHP protein, XRN2

Background
XRN2 belongs to the XRN family of 5' to 3' exoribonucleases and functions as the human ortholog of the budding yeast enzyme Rat1, operating predominantly in the nucleus to degrade RNA bearing an exposed, unprotected 5' phosphate end. XRN2 serves as the molecular torpedo in the widely established torpedo model of RNA polymerase II transcription termination: at a gene's polyadenylation signal, the endonuclease CPSF73 cleaves the nascent transcript, generating a Pol II-associated downstream RNA fragment carrying a free 5' phosphate that XRN2 loads onto directly, several bases downstream of the cleavage site, and XRN2 then degrades this fragment in the 5' to 3' direction while tracking along the transcript until it physically catches up to and dislodges the elongating polymerase. Genome-wide mapping of XRN2 substrates using a catalytically dead XRN2 mutant to trap its RNA loading sites confirms that XRN2 engages nascent RNA not only downstream of canonical polyA sites but also at the 3' ends of replication-dependent histone genes, which lack polyA tails and are instead cleaved by the same CPSF73 endonuclease, and additionally within gene bodies and near transcription start sites, where XRN2-mediated degradation of prematurely cleaved or decapped transcripts drives extensive premature termination of Pol II complexes under normal cellular conditions. Rapid, engineered depletion of XRN2 alone produces detectable transcriptional readthrough past termination sites, but this readthrough is considerably less extensive than that produced by loss of CPSF73 itself, indicating that PAS cleavage by CPSF73 provides the primary, rate-limiting step in termination while XRN2-mediated degradation accelerates and completes a process already initiated upstream. Termination efficiency is further modulated by an XRN2-linked, PP1 phosphatase-dependent mechanism in which PP1 activity slows Pol II elongation specifically over termination regions, and this polymerase slowdown facilitates XRN2's pursuit and capture of the transcribing enzyme, supporting a combined allosteric and torpedo mechanism rather than either model operating in isolation. Because XRN2-dependent termination recycles Pol II for new rounds of transcription and prevents transcriptional interference between neighboring genes, XRN2 function is directly tied to genome-wide control of gene expression boundaries, and disruption of this RNA surveillance and termination machinery is implicated in defective RNA processing relevant to cancer and to RNA-processing-linked neurodegenerative conditions.

Background

XRN2 belongs to the XRN family of 5' to 3' exoribonucleases and functions as the human ortholog of the budding yeast enzyme Rat1, operating predominantly in the nucleus to degrade RNA bearing an exposed, unprotected 5' phosphate end. XRN2 serves as the molecular torpedo in the widely established torpedo model of RNA polymerase II transcription termination: at a gene's polyadenylation signal, the endonuclease CPSF73 cleaves the nascent transcript, generating a Pol II-associated downstream RNA fragment carrying a free 5' phosphate that XRN2 loads onto directly, several bases downstream of the cleavage site, and XRN2 then degrades this fragment in the 5' to 3' direction while tracking along the transcript until it physically catches up to and dislodges the elongating polymerase. Genome-wide mapping of XRN2 substrates using a catalytically dead XRN2 mutant to trap its RNA loading sites confirms that XRN2 engages nascent RNA not only downstream of canonical polyA sites but also at the 3' ends of replication-dependent histone genes, which lack polyA tails and are instead cleaved by the same CPSF73 endonuclease, and additionally within gene bodies and near transcription start sites, where XRN2-mediated degradation of prematurely cleaved or decapped transcripts drives extensive premature termination of Pol II complexes under normal cellular conditions. Rapid, engineered depletion of XRN2 alone produces detectable transcriptional readthrough past termination sites, but this readthrough is considerably less extensive than that produced by loss of CPSF73 itself, indicating that PAS cleavage by CPSF73 provides the primary, rate-limiting step in termination while XRN2-mediated degradation accelerates and completes a process already initiated upstream. Termination efficiency is further modulated by an XRN2-linked, PP1 phosphatase-dependent mechanism in which PP1 activity slows Pol II elongation specifically over termination regions, and this polymerase slowdown facilitates XRN2's pursuit and capture of the transcribing enzyme, supporting a combined allosteric and torpedo mechanism rather than either model operating in isolation. Because XRN2-dependent termination recycles Pol II for new rounds of transcription and prevents transcriptional interference between neighboring genes, XRN2 function is directly tied to genome-wide control of gene expression boundaries, and disruption of this RNA surveillance and termination machinery is implicated in defective RNA processing relevant to cancer and to RNA-processing-linked neurodegenerative conditions.

References
https://pubmed.ncbi.nlm.nih.gov/36396340/ https://pubmed.ncbi.nlm.nih.gov/31805520/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%