AGO4 Antibody [K1B14]

Catalog No.: F4799

Application: Reactivity: Mouse, Rat, Human

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

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代表番号: 045-509-1970|電子メール：sales@selleck.co.jp

使用情報

Dilution
WB1:1000 IP1:30 IHC1:100

Application
WB, IP, IHC

Source
Rabbit Monoclonal Antibody

Reactivity
Mouse, Rat, Human

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
97 kDa 97 kDa
^*なぜ予測分子量と実際の分子量が異なるのか？下記の原因により、実際の分子量が予測と異なる：タンパク質の翻訳後修飾（リン酸化/糖鎖付加），スプライシングバリアント，イソフォーム，相対的な電荷，ポリマー。

Datasheet & SDS

生物学的記述

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

Clone
K1B14

Synonym(s)
EIF2C4; KIAA1567; AGO4; Protein argonaute-4; Argonaute4; hAgo4; Argonaute RISC catalytic component 4; Eukaryotic translation initiation factor 2C 4; eIF-2C 4; eIF2C 4

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.

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.

References
[1] Wu J, et al. J Adv Res. 2020 Apr 29;24:317-324. [2] Nakanishi K. Nucleic Acids Res. 2022 Jul 8;50(12):6618-6638.

PVDFメンブレン孔径参考表

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%