Nanotools Lc3 PDF
Contact & Orders: nanoTools Antikoerpertechnik GmbH & Co. KG Tscheulinstr. 21 . D-79331 Teningen . Germany Tel: +49-(0)7641-455 670 fax: +49-(0)7641-455 671
Aliquote and freeze in liquid nitrogen; antibody can be stored frozen at -80°C up to 1 year. Thaw aliquots at 37°C. Thawed aliquots may be stored at 4°C up to 3 months.
Stability: Aliquote and store at -20°C up to 1 year Reconstitution: liquid in PBS/0.09% Na-Azide/PEG and Sucrose/50% Glycerol (1 ml, c = 100 µg/ml)
polyclonal antibody (Rockland Immunochemicals); anti-LC3 monoclonal antibody (NanoTools Antikorpertechnik); anti-actin and anti-LAMP1 monoclonal antibodies (Sigma); and ... LC3, and ubiquitinated proteins are degraded by autophagy. Both large (more than 1 m in diameter) and small (less than
Nanotools (LC3 and 5F10), ASPP1 (LX54.2), ASPP2 (DX54.10) (5), iASPP (LX49.3), and Novocastra (p53 CM-5). Signals were detected using an ECL detection system (GE Healthcare). Autophagic Activity Assays. Autophagy was induced by several
Signaling)(1:700);LC3(NanoTools)(0.5 g/ml);AKT(Cell Signaling) (1:2000); phospho-AKT (Ser-473) (1:1000); Sirt1 (Millipore) (1:1000). GAPDH (Santa Cruz Biotechnology) (1:10,000) antibody was used as a loading control. Statistical
19Jia Wang様.mcd Page 4 09/10/30 18:53 v4.21 (M0887,Dako),Beclin-1(Sc-11427,SantaCruz,SantaCruz, CA, USA)27 and LC3 (0231s0104, Nanotools, Teningen,
AnbodiesusedweretheMitoProﬁleTotalOXPHOSkit(Mitosciences/Abcam),LC3(NanoTools)andacn(Sigma).AnalysiswasdonebynormalizingintegratedintensiestoacnlevelsusingtheOdysseysoware.StascalanalysiswasdoneusingGraphPad(Prism). ... LC3(green)andDAPI(blue).(C)Cellswerestainedwiththeautophagy
LC3 (nanoTools) or PM036 (MBL), anti-BECLIN1 (BD Biosciences), anti- ... LC3-II during the biphasic LC3-II increase (Fig. 2A), suggesting that TGF-β–mediated autophagy activation is sustained fora longer pe-riod after the early time-course activation.
173 Santa Cruz, CA) (1:500 dilution), mouse anti-LC3 (NanoTools, Teningen, Germany) 174 (1:500 dilution) or rabbit anti-GAPDH (Sigma) (1:10,000 dilution) followed by 175 extensive washing and incubation for 1 h with the appropriate horseradish
Qualitative Analysis of MAP LC3-I to LC3-II Conversion . by Western Blot . Nanotechnology Characterization Laboratory . National Cancer Institute ... 3.1.15 Mouse Monoclonal Antibody anti-LC3 antibody (NanoTools, 0231-100/LC3-5F10) 3.1.16 Peroxidase-conjugated AfiniPure Donkey Anti-Mouse ...
antibody, clone 5F10, Nanotools; 1:100 dilution following antigen retrieval) and p62/SQSTM1 (guinea pig polyclonal antibody, Progen Biotechnik; 1:100 dilution following anti- ... LC3 immunopositivity represents an early and TDP-43 immunopositivity a late marker of IBM.
and Lc3 (Nanotools 5F10 monoclonal). “*” indicates nonspecific band on ULK1 immunoblot. MeF lines were immortalized with sV40 T antigen. Below: Lc3-ii/Lc3-i signals were plotted as average ± seM, (n = 4) using 2 different y-axis scales.
The following antibodies were used for immunohistochemistry: LC3 (NanoTools), Dec1 (Giatromanolaki et al. 2003) and Ki67 (Neomarkers). The following antibodies were used for immunofluorescence; ATG12 (Cell Signaling) and HA (clone 12CA5; Cancer Research UK in house).
transferred to PVDF membranes and probed with anti-LC3 mouse polyclonal antibody (Nanotools) and anti-p62 antibody (Bethyl Laboratories). In Vivo Labeling 293T cells on 6-cm plates were transduced with plasmids and washed with
Beclin 1 Is Required for Starvation-Enhanced, but Not Rapamycin-Enhanced, LC3-Associated Phagocytosis of Burkholderia pseudomallei in RAW 264.7 Cells
Amplificationdetectionofmouseanti-LC3(0260/LC3-2G6, nanoTools),0.3%H 2 O 2 wasaddedfor10mintodestroyany residual horseradish peroxidase activity followed by a PBS wash. Cells were then incubated for 30 min in PBS-BB con-taining Triton followed by diluted rabbit anti-Atg5(FL-275,
1:1000), mouse anti-LC3 (Nanotools; 1:100), mouse anti-PML (Santa Cruz ... and LC3-positive cytoplasmic dots inATRAandATO–treated cells compared with untreated cells (Figure 3C-D). We also performed immunoﬂuorescence analysis of untreated and drug-treated cells by
LC3 (1:250; clone 2G6; Nanotools,), Atg5 (D1G9), Atg7 (all 1:500; Cell Signaling Technology) and polyclonal rabbit anti-cleaved caspase-3 (1:500) at 4°C overnight. After buffer rinse, FITC-conjugated goat anti-rabbit IgG was
1/100) from NanoTools; LC3 (polyclonal, 1/100) from MBL; slow type I MyHC (NCL-MHCS, monoclonal, 1/20), all fast type II MyHC (NCL-MHCf, monoclonal, 1/20) from Novocastra; BrdU (monoclonal, 1/200) and MEF2D (monoclonal, 1/2500) antibod-
(LC3) lipidation. Phospholipid conjugated LC3 (LC3-II) is mainly localized on autophagosomes and autolysosomes (12). ... The following primary antibodies were used: anti-LC3 (clone 5F10; Nanotools for immunofluorescence assay
(Thr389) antibodies (Cell Signaling Technologies); monoclonal anti-LC3 antibody(nanoTools,thoughAxxora;LLC). siRNApreparationandtransfection.siRNAsequencetargetingeEF-2 kinase mRNA corresponded to the coding region 144 to 164 (5¶- ... formation of LC3-II, but also triggered apoptosis in a dose-
dependent on LC3 abundance, suggesting that TBC1D5 can switch from retromer-decorated endosomal to LC3-lipidated ... (NanoTools), rabbit polyclonal anti-LC3 (MBL) and rabbit polyclonal anti-LC3B (a gift from Z. Elazar), rabbit polyclonal anti-p62 (Enzo Life
anti-LC3, and anti- -tubulin ... (Cambridge, MA), nanoTools (Teningen, Germany), and Sigma-Aldrich, respectively. Anti-WIPI-1 antiserum was described previously (Proikas-Cezanne et al., 2004). Anti-rabbit IgG-Alexa Fluor 546 and TO-PRO-3 was obtained from Invitrogen.
antibody for the autophagy marker, LC3 (LC3-5F10; 30 g/mouse; NanoTools, San Diego, CA, USA) was injected into 3 mice in each of the 4 experimental groups to label the autophagic osteocytes. All animals were treated accord-
clonal anti–LC3 antibody (1:100; Nanotools Antik€orper-technik GmbH & Co Antik€orpertechnik GmbH & Co; clone 5F10) as primary antibodies. Incubation with Alexa-488–conjugated secondary antibody (Invitrogen) was done in the dark at room temperature for 1 hour.
mouse α-LC3 (1:1000, NanoTools 0260), rabbit α-β-tubulin (1:20000, Santa Cruz, sc-9104), ... LC3-II levels due to the accumulation of undigested autophagosomes. p62 is a protein degraded by au-tophagy and commonly monitored as a marker of
Biotechnology); and anti-LC3 (Nanotools, Munich, Germany). Immunohistochemistry Immunohistochemical analyses were performed as described previ-ously.14,18 Primary antibodies were purchased as follows: anti-HA, anti-Lamp1, and anti-cathepsin D (Santa Cruz Biotechnology);
Adescribedα-LC3 rabbitpolyclonalsera was used for immunoblotting (5). ... 5F10;NanoTools/Axorra)for1hatroomtemperature.Secondary antibody (OmniMap anti-Ms HRP; Ventana) was applied for 12 min, and ChromoMap DAB (Ventana) was used for detection.
Transient DsRed–LC3 and green fluorescent protein–Endo transfection. PCR-amplified coding sequences of LC3 in green fluorescent protein (GFP) ... (1:1000; TUJ-1; R&D Systems), overnight for LC3 (1:250; Nanotools) and cathepsin D (1:5000; in house) at 4°C. Cells were washed (three
Staining of endogenous LC3 : 12 µm-thick liver and lung sect ions of paraffin-embedded WT ... (clone 5F10 from Nanotools, Teningen, Germany) overnight. Sections were then incubated with FITC-streptavidin (1:100) (Vector Laboratories, INC.,
Mouse anti-LC3 antibody (NanoTools, Antikörpertechnik GmbH & Co, Teningen, Germany). 2. pBABE-GFP-LC3 vector (Dr. J. Debnath, Dept. Cell Biol- ... LC3-I is highly susceptible to degradation in the lysis buffer, especially during freeze–thaw cycles.
against LC3 (Clone 5F10, Nanotools), followed by Alexa488-labeled donkey anti-mouse IgG (Molecular Probes, Life Technologies). ... Quantification of LC3 spots was performed by the spot-detection protocol in the PhysiologyAnalyst
Signaling Technology, Danvers, MA), LC3 (Nanotools, Teningen, Germany), and LRRK2 (C41-2; Michael J. Fox Foundation, New York City, NY). Calcium Imaging ... LC3 suppresses neurite shortening caused by mutant LRRK2.12,19 However, the substrate(s) removed by auto-
clonal antibody to LC3 (1:1200; Nanotools). Secondary antibodies were HRP-conjugated goat anti-mouse, goat anti-rabbit IgG (1:10,000; Pro- ... GFP–LC3 cDNA construct (Schmid et al., 2007) was transfected into HeLa cells grown on coverslips, ...
77 (Lorrach, Germany); anti-LC3 (clone 5F10) from NanoTools (Teningen, Germany); and 78 anti-β-actin from Sigma-Aldrich. Secondary antibodies HRP donkey anti-rabbit and 79 sheep anti-mouse were purchased from GE Healthcare (Bucks, UK). 80 81 ...
Nanotools の抗体と両方 ... TP53INP1, a tumor suppressor, interacts with LC3 and ATG8 family proteins through the LC3interacting region (LIR) and promotes autophagydependent cell death 22421968 2012 Cell Death Differ in press IC Human, Mouse U2OS, HeLa.
Mouse anti-LC3 antibody (NanoTools, Antikörpertechnik GmbH&Co,Teningen,Germany). 2. pBABE-GFP-LC3vector(Dr.J.Debnath,Dept.CellBiol- ... LC3 have been raised in a different species and that the correspondingsecondaryantibodiesarelabeledwithdiffer-
147 ß-actin was from Sigma (Munich, Germany), mouse-monoclonal anti-LC3 antibody was 148 obtained from nanoTools Antikörpertechnik GmbH & Co., KG (Teningen, Germany) and the 149 mouse monoclonal anti-ACE2 antibody was obtained from Enzo Life Science (Lörrach,
Material y métodos - 174 - Belgium). El anticuerpo frente LC3 (clone 5F10) es de Nanotools (Teningen, Germany), el anticuerpo frente al TGN46 humano de Serotec (Oxford, UK) y el anticuerpo ATP6J frente a