Datasheet

生物学的記述

Specificity	AGO4 Antibody [K1B14] detects endogenous levels of total AGO4 protein.
Background	AGO4 (Argonaute RISC Catalytic Component 4), a member of the mammalian Argonaute family specialized in siRNA-guided gene silencing and heterochromatin formation, adopts a bilobed architecture with an N-terminal PAZ domain anchoring the 3′ end of guide siRNAs via a conserved OB-fold pocket, a central PIWI (slicer-incompetent) domain featuring an AGO4-specific insertion (4SI, E629-Q638) protruding into the nucleic acid-binding channel on Loop 2 to modulate target accessibility, and MID domain clamping the guide 5′ phosphate, connected by flexible linkers enabling open-closed conformational transitions. Loaded with 21-24 nt siRNAs or miRNAs via Dicer/TRBP handover, AGO4 assembles functional RISC through PAZ-mediated passenger strand ejection and PIWI-driven guide positioning where seed nucleotides (g2-g8) form A-form helix seed-pairing with target mRNA, inducing translational repression, deadenylation, or decapping via GW182 recruitment despite lacking slicer activity due to mutated catalytic Asp/Glu/His triad (replaced by Gly/Arg), while uniquely directing DNA methylation at promoter-proximal loci through TNRC6A/DNMT3A interactions promoting H3K9me3 and silencing tumor suppressors or retrotransposons. Predominantly cytoplasmic but shuttling nuclearly, AGO4 underpins heterochromatin maintenance at pericentromeric repeats and developmental gene repression, with dysregulation implicated in hepatocellular carcinoma via miR-3191-5p-mediated IRES inhibition and global hypomethylation fostering chromosomal instability.

Specificity

AGO4 Antibody [K1B14] detects endogenous levels of total AGO4 protein.

Background

AGO4 (Argonaute RISC Catalytic Component 4), a member of the mammalian Argonaute family specialized in siRNA-guided gene silencing and heterochromatin formation, adopts a bilobed architecture with an N-terminal PAZ domain anchoring the 3′ end of guide siRNAs via a conserved OB-fold pocket, a central PIWI (slicer-incompetent) domain featuring an AGO4-specific insertion (4SI, E629-Q638) protruding into the nucleic acid-binding channel on Loop 2 to modulate target accessibility, and MID domain clamping the guide 5′ phosphate, connected by flexible linkers enabling open-closed conformational transitions. Loaded with 21-24 nt siRNAs or miRNAs via Dicer/TRBP handover, AGO4 assembles functional RISC through PAZ-mediated passenger strand ejection and PIWI-driven guide positioning where seed nucleotides (g2-g8) form A-form helix seed-pairing with target mRNA, inducing translational repression, deadenylation, or decapping via GW182 recruitment despite lacking slicer activity due to mutated catalytic Asp/Glu/His triad (replaced by Gly/Arg), while uniquely directing DNA methylation at promoter-proximal loci through TNRC6A/DNMT3A interactions promoting H3K9me3 and silencing tumor suppressors or retrotransposons. Predominantly cytoplasmic but shuttling nuclearly, AGO4 underpins heterochromatin maintenance at pericentromeric repeats and developmental gene repression, with dysregulation implicated in hepatocellular carcinoma via miR-3191-5p-mediated IRES inhibition and global hypomethylation fostering chromosomal instability.

使用情報

Application

WB, IP, IHC

Dilution

WB	IP	IHC
1:1000	1:30	1:100

Reactivity

Mouse, Rat, Human

Source

Rabbit Monoclonal Antibody

MW

97 kDa

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

プロトコール

References

https://pubmed.ncbi.nlm.nih.gov/32455006/
https://pubmed.ncbi.nlm.nih.gov/35736234/

Application Data

WB

Validated by Selleck

Lane 1: Hela, Lane 2: THP-1, Lane 3: 3T3, Lane 4: C6