ATG4B Antibody [P14M3]

Catalog No.: F4916

Application: Reactivity: Mouse, Human

Lane 1: Ramos, Lane 2: 293T, Lane 3: Jurkat, Lane 4: Hela

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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 - 1:10000 FCM1:100 - 1:500

Application
WB, FCM

Source
Rabbit Monoclonal Antibody

Reactivity
Mouse, 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
44 kDa 47 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
ATG4B Antibody [P14M3] detects endogenous levels of total ATG4B protein.

Clone
P14M3

Synonym(s)
APG4B, AUTL1, KIAA0943, ATG4B, Cysteine protease ATG4B, AUT-like 1 cysteine endopeptidase, Autophagy-related cysteine endopeptidase 1, Autophagy-related protein 4 homolog B, Autophagin-1, HsAPG4B, Hapg4b

Background
ATG4B is a crucial cysteine protease in autophagy, responsible for processing and recycling ATG8 family proteins such as LC3 and GABARAP, thereby enabling autophagosome biogenesis across eukaryotes. ATG4B contains a catalytic triad composed of Cys74, Asp278, and His280 within a papain-like fold, with a regulatory loop and Trp142 that maintain the enzyme in an autoinhibited state until substrate engagement induces conformational changes for peptide bond hydrolysis. N-terminal motifs further enhance specificity for LC3 conjugated to phosphatidylethanolamine (PE) on autophagic membranes. ATG4B’s primary function is to expose the C-terminal glycine on pro-LC3 through an initial priming cleavage, preparing it for conjugation to PE via the ATG7/ATG3 ubiquitin-like cascade, and subsequently to catalyze the delipidation of LC3-PE on the outer autophagosomal membrane, facilitating the recycling of free LC3. With the broadest substrate selectivity among ATG4 paralogs, ATG4B primarily favors LC3, balancing autophagosome membrane elongation, tethering, and autophagic flux. Its activity is tightly regulated through redox modifications at Cys292 and Cys361 and by phosphorylation, adapting autophagy to cellular stress; overexpression of wild-type ATG4B can block LC3 lipidation by excessive deconjugation, while catalytically inactive mutants like C74A sequester LC3 and inhibit autophagy. Dysregulation of ATG4B activity is linked to tumorigenesis due to impaired autophagic flux.

Background

ATG4B is a crucial cysteine protease in autophagy, responsible for processing and recycling ATG8 family proteins such as LC3 and GABARAP, thereby enabling autophagosome biogenesis across eukaryotes. ATG4B contains a catalytic triad composed of Cys74, Asp278, and His280 within a papain-like fold, with a regulatory loop and Trp142 that maintain the enzyme in an autoinhibited state until substrate engagement induces conformational changes for peptide bond hydrolysis. N-terminal motifs further enhance specificity for LC3 conjugated to phosphatidylethanolamine (PE) on autophagic membranes. ATG4B’s primary function is to expose the C-terminal glycine on pro-LC3 through an initial priming cleavage, preparing it for conjugation to PE via the ATG7/ATG3 ubiquitin-like cascade, and subsequently to catalyze the delipidation of LC3-PE on the outer autophagosomal membrane, facilitating the recycling of free LC3. With the broadest substrate selectivity among ATG4 paralogs, ATG4B primarily favors LC3, balancing autophagosome membrane elongation, tethering, and autophagic flux. Its activity is tightly regulated through redox modifications at Cys292 and Cys361 and by phosphorylation, adapting autophagy to cellular stress; overexpression of wild-type ATG4B can block LC3 lipidation by excessive deconjugation, while catalytically inactive mutants like C74A sequester LC3 and inhibit autophagy. Dysregulation of ATG4B activity is linked to tumorigenesis due to impaired autophagic flux.

References
[1] Maruyama T, et al. J Antibiot (Tokyo). 2017 Sep 13;71(1):72–8. [2] Park NY, et al. Cells. 2022 Apr 13;11(8):1330.

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%