Phospho-Atg14 (Ser29) Antibody [B12C19]

Application: Reactivity: Human, Mouse, Rat

Lane 1: Saos-2, Lane 2: Saos-2 (EBSS, 2 h)

Usage Information

Dilution
WB1:1000 IF1:400-1:800 FCM1:400-1:1600

Application
WB, 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
65 kDa

Datasheet & SDS

Biological Description

Specificity
Phospho-Atg14 (Ser29) Antibody [B12C19] detects endogenous levels of total Atg14 protein only when it is phosphorylated at Ser29.

Clone
B12C19

Synonym(s)
Beclin 1-associated autophagy-related key regulator; Barkor; Autophagy-related protein 14-like protein; Atg14L; ATG14; ATG14L; KIAA0831

Background
Phospho-Atg14 (Ser29) designates a key regulatory phosphorylation on Atg14L, a core subunit of the PI3KC3 (Vps34) complex pivotal to autophagy initiation, where it associates with Beclin-1, Vps34, and Vps15 to generate PI3P at nascent phagophores, driving membrane curvature and LC3 lipidation for autophagosome biogenesis. Atg14L features a coiled-coil domain for Beclin-1 heterodimerization and a BATS domain that anchors the complex to ER subdomains and isolation membranes, positioning Ser29 within an N-terminal regulatory motif sensitive to kinase modulation. TBK1 phosphorylates Ser29 in response to nutrient stress or pathogen sensing, enhancing Atg14L recruitment to omegasomes and boosting Vps34 lipid kinase activity through allosteric conformational shifts that promote PI3P production and Atg9 trafficking for phagophore expansion. This modification integrates into the ULK1-TBK1-Beclin-1 signaling axis, where Ser29 phosphorylation synergizes with ULK1-mediated Beclin-1 activation to amplify autophagy flux under starvation, while counteracting mTORC1 suppression ensures rapid autophagosome formation for selective cargo engulfment like damaged mitochondria. Phospho-Ser29 drives dynamic membrane remodeling in diverse cellular contexts, from neuronal survival during ER stress to immune cell activation against intracellular pathogens, making it invaluable for researchers tracking autophagy kinetics via phospho-specific probes or dissecting TBK1 dependency in high-throughput screens. Dysregulation through hypo-phosphorylation impairs autophagic clearance, contributing to neurodegeneration and tumorigenesis, whereas hyperactivation risks excessive catabolism in metabolic disorders.

Background

Phospho-Atg14 (Ser29) designates a key regulatory phosphorylation on Atg14L, a core subunit of the PI3KC3 (Vps34) complex pivotal to autophagy initiation, where it associates with Beclin-1, Vps34, and Vps15 to generate PI3P at nascent phagophores, driving membrane curvature and LC3 lipidation for autophagosome biogenesis. Atg14L features a coiled-coil domain for Beclin-1 heterodimerization and a BATS domain that anchors the complex to ER subdomains and isolation membranes, positioning Ser29 within an N-terminal regulatory motif sensitive to kinase modulation. TBK1 phosphorylates Ser29 in response to nutrient stress or pathogen sensing, enhancing Atg14L recruitment to omegasomes and boosting Vps34 lipid kinase activity through allosteric conformational shifts that promote PI3P production and Atg9 trafficking for phagophore expansion. This modification integrates into the ULK1-TBK1-Beclin-1 signaling axis, where Ser29 phosphorylation synergizes with ULK1-mediated Beclin-1 activation to amplify autophagy flux under starvation, while counteracting mTORC1 suppression ensures rapid autophagosome formation for selective cargo engulfment like damaged mitochondria. Phospho-Ser29 drives dynamic membrane remodeling in diverse cellular contexts, from neuronal survival during ER stress to immune cell activation against intracellular pathogens, making it invaluable for researchers tracking autophagy kinetics via phospho-specific probes or dissecting TBK1 dependency in high-throughput screens. Dysregulation through hypo-phosphorylation impairs autophagic clearance, contributing to neurodegeneration and tumorigenesis, whereas hyperactivation risks excessive catabolism in metabolic disorders.

References
https://pubmed.ncbi.nlm.nih.gov/27938392/ https://pubmed.ncbi.nlm.nih.gov/40442316/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

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%