ATG13 phospho S318 Antibody, Rabbit, Polyclonal

Artikelname:	ATG13 phospho S318 Antibody, Rabbit, Polyclonal
Artikelnummer:	ROC-600-401-C49S
Hersteller Artikelnummer:	600-401-C49S
Alternativnummer:	ROC-600-401-C49S
Hersteller:	Rockland Immunochemicals
Wirt:	Rabbit
Kategorie:	Antikörper
Applikation:	DOT, ELISA, WB
Spezies Reaktivität:	Human
Immunogen:	This affinity purified antibody was prepared by repeated immunizations with a synthetic peptide corresponding to the region near S318 of ATG13.
Konjugation:	Unconjugated
Alternative Synonym:	rabbit anti-ATG13 pS318 Antibody, ATG-13, ATG 13, Autophagy-related protein 13, KIAA0652

Klonalität:	Polyclonal
Konzentration:	1.1 mg/mL by UV absorbance at 280 nm
NCBI:	9776
UniProt:	O75143
Puffer:	0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Formulierung:	Liquid (sterile filtered)
Target-Kategorie:	Human
Antibody Type:	Primary Antibody

Application Verdünnung:	ELISA: 1:25,000-1:175,000, Flow Cytometry: User Optimized, WB: 1:1000
Anwendungsbeschreibung:	This affinity purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 56.6 kDa in size corresponding to human phosphorylated ATG13 protein



	Western Blot of Rabbit Anti-ATG13pS318 Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: BSA Conjugated ATG13 phospho S318 peptide - reduced [0.2µg]. Lane 3: BSA Conjugated ATG13 non-phospho S318 peptide - reduced [0.2µg]. Primary Antibody: Anti-ATG13pS318 [Rabbit] Antibody at 1.0µg/mL overnight at 2-8C. Secondary Antibody: Anti-Rabbit IgG [Goat] Peroxidase conjugated (p/n 611-1302) at 1:40,000 for 30mins at RT. Block: Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) for 1hr at RT. Expected: detects the phospho S318 peptide and does not detect the NP-peptide.Exposure: 6.1sec.
	Western blot using Rocklands affinity purified anti-ATG13 pS318 antibody shows detection of phosphorylated ATG13 in 293T cells engineered to coexpress Ulk1 and Atg13 (Ulk1 + Atg13). In the left lane was loaded kinase-dead hypophosphorylated Ulk1-K46A mutant + ATG13. The right lane contains the 293T Ulk1 + ATG13 lysate and shows detection at approximately 57 kDa. The antibody was purified and resolved by SDS-PAGE, then transferred to nitrocellulose membrane. The membrane was blocked with 5% Blotto (p/n B501-0500) and probed with the primary antibody at 1µg/mL overnight at 4C. After washing, the membrane was probed with Goat Anti-Rabbit HRP secondary 1:5000 in detection buffer (p/n MB-070) for 45 minutes at room temperature. In collaboration with Charles Dorsey at Eli Lilly, Indianapolis, IN and John Cleveland at Scripps, Jupiter, FL.
	Autophagic vesicles are locally formed in dendrites of cultured neurons following LTD. A) Top, representative super-resolution microscopy dSTORM image of a secondary dendrite labeled with an antibody against LC3, 15min after cLTD. Bottom, magnification of representative U-shaped LC3-positive structures in dendrites, 15min after NMDA or DHPG pulses. Scale bars: 2µm and 250nm, as indicated. (N=3 independent experiments). B) Graph showing the number of LC3-positive U-shaped structures in secondary dendrites visualized in (a), before (control) and 15min or 60min after NMDAR- and mGluR-LTD. Bars represent mean valuesSEM. N=3 independent experiments per condition (n>9 dendrites per condition). Statistical analysis was performed by one-way ANOVA. For the time point of 15minF(2, 33)=17.93, p<0.0001) (Tukeys test Pcontrol-NMDA<0.0001, Pcontrol-DHPG<0.0001). For the time point of 60minF(2, 21)=9.459, p=0.0012) (Tukeys test Pcontrol-NMDA=0.0041, Pcontrol-DHPG=0.0024). C) Graph showing the size distribution (nm) of the dendritic U-shaped LC3-positive structures visualized in a upon NMDAR- and mGluR-LTD. Bars represent mean valuesSEM for each analysed dendrite. N=3 independent experiments (n>40 dendrites per condition). D) Confocal images of dendrites immunolabeled with antibodies against WIPI2, LC3, and MAP2 before (control) or after 15min of NMDAR- and mGluR-LTD. Scale bar: 10µm. (N=6 independent experiments). E) Representative confocal images of neurons immunolabeled with antibodies against ULK1, Atg101, Atg13, FIP200 and, along with MAP2 to label dendrites before (control) or 15min after LTD-inducing pulses. Scale bar: 20µm. Graphs showing the number of puncta positive for each ULK1-complex component in secondary dendrites, normalized for dendrite length, in every condition, as indicated. Graph bars represent mean valuesSEM. N=6 independent experiments per condition. Statistical analyses were performed using one-way ANOVA. ULK1: F(2,15)=24.48, P<0.0001 (Tukeys multiple comparison test, Pcontrol_NMDAR<0.0001, Pcontrol_mGluR<0.0001, PNMDAR_mGluR=0.8825). Atg101: F(2,15)=24.31, P<0.0001 (Tukeys multiple comparison test, Pcontrol_NMDAR<0.0001, Pcontrol_mGluR<0.0001, PNMDAR_mGluR=0.9329). Atg13: F(2,15)=8.386, P=0.0036 (Tukeys multiple comparison test, Pcontrol_NMDAR=0.007, Pcontrol_mGluR=0.0086, PNMDAR_mGluR=0.9940). FIP200: F(2,15)=17.66, P=0.0001 (Tukeys multiple comparison test, Pcon