ATG5 Antibody

Western Blot: ATG5 Antibody [NB110-53818] - Analysis in mouse wildtype ES cell lysate (Lane 1) using NB110-53818. Lane 2 is a mouse ATG5 KO ES cell lysate (negative control). Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell Biol. 152 (2001)] Photo courtesy of Dr. Beth Levine, UT Southwestern Medical Center.

Western Blot: ATG5 Antibody [NB110-53818] - Total protein from Human HeLa and A431 and Mouse MEF cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-ATG5 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunocytochemistry/Immunofluorescence: ATG5 Antibody [NB110-53818] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-ATG5 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: ATG5 Antibody [NB110-53818] - Staining of human liver hepatocytes at 2.5ug/ml. 40X magnification.

Immunocytochemistry/Immunofluorescence: ATG5 Antibody [NB110-53818] - Staining of SY5Y cells at 1:250. Incubated overnight at 4 degrees. Photo courtesy of an anonymous collaborator.

Immunocytochemistry/Immunofluorescence: ATG5 Antibody [NB110-53818] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ATG5 [NB110-53818] at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: ATG5 Antibody [NB110-53818] - Staining mouse intestine using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: ATG5 Antibody [NB110-53818] - Staining of mouse knee. Image from verified customer review.

Simple Western: ATG5 Antibody [NB110-53818] - Simple Western lane view shows a specific band for ATG5 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Po, Bv, Pm, Xp, Ze
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP, PLA, RIA, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

ATG5 Antibody Summary

• Immunogen: A synthetic peptide made to an N-terminal region of the human ATG5 protein (within residues 1-50) [Swiss-Prot Q9H1Y0]
• Localization: Cytoplasm. Co-localizes with non-muscle actin.
• Specificity: This is selective for the full-length and calpain cleaved isoform proteins. The short isoform is missing amino acids 1-79. The calpain cleaved form of ATG5 is missing amino acids 195-275.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ATG5
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:500
  • Simple Western 1:50
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:250
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Paraffin 1:400
  • Immunoprecipitation 1:10 - 1:500
  • Proximity Ligation Assay
  • Radioimmunoassay
  • Knockout Validated
• Application Notes: In Western Blot, a band is seen ~56 kDa representing the ATG5-ATG12 complex, the molecular weight of human ATG5 is ~33 kDa. In ICC/IF, cytoplasmic staining was observed in SY5Y cells. In IHC-P, staining was observed in the cytoplasm of human hepatocytes and mouse intestine tissues. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Use in Immunoprecipitation reported in scientific literature (PMID 24705551). Use in radioimmunoassay reported in scientific literature (PMID: 28651493). PLA data from independent lab. Use in electron microscopy reported in scientific literature (PMID 27219062).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Control:

  ATG5 Lysate (NBL1-07799)

• Control Peptide:

  ATG5 Peptide (NB110-53818PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Purity: Immunogen affinity purified

Alternate Names for ATG5 Antibody

• APG5 autophagy 5-like (S. cerevisiae)
• APG5
• APG5LS. cerevisiae)-like
• ASP
• ASPAPG5-LIKE
• ATG5 autophagy related 5 homolog (S. cerevisiae)
• ATG5
• Autophagy protein 5

Background

ATG5 (autophagy protein 5, also known as APG5-like, apoptosis-specific protein, APG5L, and ASP) is an essential protein for autophagy. It is expressed ubiquitously, and expression increases dramatically in apoptotic cells. ATG5 is localized in the cytoplasm (can colocalize with nonmuscle actin) and is induced upon apoptotic stimuli. ATG5-ATG12 can form a complex with several units of ATG16, inhibiting ATG5-TECPR1 direct interactions. After joining to ATG12, ATG5 associates with the isolation membrane to form autophagosomes, mediated by ubiquitin E1-like enzyme Atg7 as well as the E2-like enzyme Atg10. ATG5 also plays an important role in the apoptotic process within the modified cytoskeleton, and its expression is a relatively late event in apoptotic process, occurring downstream of caspase activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ATG5 Antibody (NB110-53818)(157)

Publications using NB110-53818

• Yao J, Qiu Y, Frontera E et al. Inhibiting autophagy reduces retinal degeneration caused by protein misfolding. Autophagy. Jul 13 2018 [PMID: 29940785] (WB, Mouse)
• Gibson J. In vivo imaging and analysis of host-pathogen interactions of intracellular pathogens Thesis 2017 (WB, Zebrafish)
• Karanasios E, Ktistakis NT. Studying Autophagy: List of Useful Antibodies Produced via a Community Effort. Autophagy at the Cell, Tissue and Organismal Level. 2016 Apr 24
• Reed M. Requirement of autophagy proteins Beclin-1 and Map1-LC3b in the induction of innate and adaptive immune responses to respiratory syncytial virus infection. Thesis. 2014 (ICC/IF, Mouse)
• Wan W, You Z, Zhou L et al. mTORC1-Regulated and HUWE1-Mediated WIPI2 Degradation Controls Autophagy Flux. Mol. Cell. Oct 18 2018 [PMID: 30340022] (WB)
• Gorgulu k, Diakopoulos kN, Ai J et al. Levels of the Autophagy Related 5 Protein Affect Progression and Metastasis of Pancreatic Tumors in Mice. Gastroenterology. Oct 5 2018 [PMID: 30296435] (IHC, Mouse)
• Ye T, Jiang K, Wei L et al. Oncolytic Newcastle disease virus induces autophagy-dependent immunogenic cell death in lung cancer cells. Am J Cancer Res Aug 1 2018 [PMID: 30210920] (WB, Human)
• Wu XM, Chen WQ, Hu YW et al. RIP2 Is a Critical Regulator for NLRs Signaling and MHC Antigen Presentation but Not for MAPK and PI3K/Akt Pathways. Front Immunol Apr 10 2018 [PMID: 29692779] (WB, Zebrafish)
• Murera D, Arbogast F, Arnold J et al. CD4 T cell autophagy is integral to memory maintenance. Sci Rep Apr 13 2018 [PMID: 29654322] (WB, Mouse)
• Kim YH, Kwak MS, Shin JM et al. Inflachromene inhibits autophagy through modulation of Beclin 1 activity. J Cell Sci 2018 Feb 20 [PMID: 29361549]
• Xia X, Wang L, Zhang X et al. Halofuginone-induced Autophagy Suppresses the Migration and Invasion of MCF-7 Cells via Regulation of STMN1 and p53. J. Cell. Biochem. 2017 Dec 12 [PMID: 29231257] (Human)
• Byun YS, Lee HJ, Shin S, Chung SH. Elevation of autophagy markers in Sjogren syndrome dry eye. Sci Rep 2017 Dec 08 [PMID: 29222450] (WB, Human)
• Yang PW, Hsieh MS, Chang YH et al. Genetic polymorphisms of ATG5 predict survival and recurrence in patients with early-stage esophageal squamous cell carcinoma. Oncotarget 2017 Oct 31 [PMID: 29207660] (Human)
• Baron O, Boudi A, Dias C et al. Stall in Canonical Autophagy-Lysosome Pathways Prompts Nucleophagy-Based Nuclear Breakdown in Neurodegeneration Curr. Biol. 2017 Nov 17 [PMID: 29174892] (Mouse)
• Sun P, Zhang S, Qin X et al. Foot-and-mouth disease virus capsid protein VP2 activates the cellular EIF2S1-ATF4 pathway and induces autophagy via HSPB1 Autophagy. 2017 Nov 23 [PMID: 29166823] (Bovine)
• Martinez-Lopez N, Tarabra E, Toledo M et al. System-wide Benefits of Intermeal Fasting by Autophagy. Cell Metabolism. [PMID: 29107505] (Mouse)
• Fiorito F, Iovane V, Cantiello A et al. MG-132 reduces virus release in Bovine herpesvirus-1 infection. Sci Rep. 2017 Oct 17 [PMID: 29042667] (WB, Bovine)
• Kuss-Duerkop SK, Wang J, Mena I et al. Influenza virus differentially activates mTORC1 and mTORC2 signaling to maximize late stage replication. PLoS Pathog. 2017 Sep 01 [PMID: 28953980] (WB, Human)
• Reed M. Requirement of autophagy proteins Beclin-1 and Map1-LC3b in the induction of innate and adaptive immune responses to respiratory syncytial virus infection Thesis 2014 (ICC/IF, Mouse)
  
  Details:
  This reference used the DyLight 550 version of NB110-53818.
• Meckes JK, Carames B, Olmer M et al. Compromised autophagy precedes meniscus degeneration and cartilage damage in mice Osteoarthr. Cartil. 2017 Aug 08 [PMID: 28801209] (IHC-P, Mouse)
• Mans LA, Querol Cano L, van Pelt J et al. The tumor suppressor LKB1 regulates starvation-induced autophagy under systemic metabolic stress Sci Rep 2017 Aug 04 [PMID: 28779098] (WB, Zebrafish)
• Reed M. Requirement of autophagy proteins Beclin-1 and Map1-LC3b in the induction of innate and adaptive immune responses to respiratory syncytial virus infection. Thesis. 2014 (ICC/IF, Mouse)
  
  Details:
  Citation using the DyLight 550 form of this antibody.
• Xia X, Gao Y, Zhang J et al. Autophagy mediated by arginine depletion activation of the nutrient sensor GCN2 contributes to interferon-gamma-induced malignant transformation of primary bovine mammary epithelial cells Cell Death Discovery Jan 25 2016 (WB, Bovine)
• Zhang R, Xu J, Zhao J, Bai, J. Mir-30d suppresses cell proliferation of colon cancer cells by inhibiting cell autophagy and promoting cell apoptosis. Tumour Biol. 2017 Jun 01 [PMID: 28651493] (WB, Flow, RIA, Human)
• Li Z, Yuan Y, Meng Y et al. Autophagy upregulation ameliorates cell injury in Sequestosome 1 knockout podocytes in vitro. Biochem. Biophys. Res. Commun. 2017 Jun 07 [PMID: 28600173]
• Hine C, Kim HJ, Zhu Y et al. Hypothalamic-Pituitary Axis Regulates Hydrogen Sulfide Production Cell Metab. Jun 6 2017 [PMID: 28591635] (WB, Mouse)
• Zielniok K, Sobolewska A, Gajewska M. Mechanisms of autophagy induction by sex steroids in bovine mammary epithelial cells. J. Mol. Endocrinol. Apr 20 2017 [PMID: 28428343]
• Okerlund ND, Schneider K, Leal-Ortiz S et al. Bassoon Controls Presynaptic Autophagy through Atg5. Neuron. 2017 Feb 22 [PMID: 28231469]
• Maus M, Cuk M, Patel B et al. Store-Operated Ca2+ Entry Controls Induction of Lipolysis and the Transcriptional Reprogramming to Lipid Metabolism. Cell Metab. Mar 7 2017 [PMID: 28132808] (WB, Human)
• Fabrice Gankam-Kengne, Bruno S. Couturier, Alain Soupart et al. Osmotic Stress-Induced Defective Glial Proteostasis Contributes to Brain Demyelination after Hyponatremia Treatment. jasn.org 2017 Jan 25 [PMID: 28122966] (WB, Rat)
• Cai Y, Yang L, Hu G et al. Regulation of morphine-induced synaptic alterations: Role of oxidative stress, ER stress, and autophagy. J. Cell Biol. Oct 24 2016 [PMID: 27810915] (WB, Rat)
• Starobinets H, Ye J, Broz M et al. Antitumor adaptive immunity remains intact following inhibition of autophagy and antimalarial treatment. J. Clin. Invest. Oct 24 2016 [PMID: 27775547]
• Lee E, Wei Y, Zou Z et al. Genetic inhibition of autophagy promotes p53 loss-of-heterozygosity and tumorigenesis. Oncotarget. Oct 18 2016 [PMID: 27655644] (WB, Zebrafish)
• Tan X, Wang Hc, Liang Ry, Chuang Sm. Human Rad23A plays a regulatory role in autophagy. Biochem. Biophys. Res. Commun. Sep 30 2016 [PMID: 27613096]
• Sousa CM, Biancur DE, Wang X et al. Pancreatic stellate cells support tumour metabolism through autophagic alanine secretion. Nature. Aug 25 2016 [PMID: 27509858] (WB)
• Choi MS, Kim HJ, Ham M et al. Amber Light (590 nm) Induces the Breakdown of Lipid Droplets through Autophagy-Related Lysosomal Degradation in Differentiated Adipocytes. Sci Rep 2016 Jun 27 [PMID: 27346059] (WB)
• Goodall ML, Fitzwalter BE, Zahedi S et al. The Autophagy Machinery Controls Cell Death Switching between Apoptosis and Necroptosis. Dev. Cell. May 23 2016 [PMID: 27219062] (PLA, EM, Mouse)
• Orhon I, Dupont N, Zaidan M et al. Primary-cilium-dependent autophagy controls epithelial cell volume in response to fluid flow. Nat. Cell Biol. Jun 1 2016 [PMID: 27214279] (WB, Mouse)
• Sharifi MN, Mowers EE, Drake LE et al. Autophagy Promotes Focal Adhesion Disassembly and Cell Motility of Metastatic Tumor Cells through the Direct Interaction of Paxillin with LC3. Cell Rep. May 24 2016 [PMID: 27184837] (WB, Mouse)
• Song JX, Sun YR, Peluso I et al. A novel curcumin analog binds to and activates TFEB in vitro and in vivo independent of MTOR inhibition. Autophagy. May 12 2016 [PMID: 27172265] (WB, Mouse)
• Choudhury KR, Bucha S, Baksi S et al. Chaperone-like protein HYPK and its interacting partners augment autophagy. Eur. J. Cell Biol. 2016 Apr 01 [PMID: 27067261] (WB, Mouse)
• Xia X, Che Y, Gao Y et al. Arginine Supplementation Recovered the IFN-gamma-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2alpha Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells. Mol Cells 2016 May 31 [PMID: 27025389] (WB)
• Yun Woo Jin, Kim Eun-Young, Park Ji-Eun et al. Microtubule-associated protein light chain 3 is involved in melanogenesis via regulation of MITF expression in melanocytes. Scientific Reports 2016 [PMID: 26814135] (WB, Mouse)
• Imam S, Talley S, Nelson RS et al. TRIM5alpha degradation via autophagy is not required for retroviral restriction. J. Virol. 2016 Jan 13 [PMID: 26764007] (WB, Human)
• Martinez-Lopez N, Garcia-Macia M, Sahu S. Autophagy in the CNS and Periphery Coordinate Lipophagy and Lipolysis in the Brown Adipose Tissue and Liver. Cell Metabolism. 2015 Nov 19 [PMID: 26698918] (WB, Mouse)
• Zhang F, Wang J, Chu J et al. MicroRNA-146a Induced by Hypoxia Promotes Chondrocyte Autophagy through Bcl-2. Cell. Physiol. Biochem. 2015 Nov 06 [PMID: 26492575] (WB, Mouse)
  
  Details:
  ATG5 antibody used for WB on isolated cells from mouse hip and knee joints at a dilution of 1:500 (Figure 3E).
• Baisantry A, Bhayana S, Rong S et al. Autophagy Induces Prosenescent Changes in Proximal Tubular S3 Segments. J. Am. Soc. Nephrol. Jun 1 2016 [PMID: 26487561] (IHC-P, Mouse)
• Liu L, Li CJ, Lu Y et al. Baclofen mediates neuroprotection on hippocampal CA1 pyramidal cells through the regulation of autophagy under chronic cerebral hypoperfusion. Sci Rep 2015 [PMID: 26412641] (WB)
• Pal I, Parida S, Prashanth Kumar BN et al. Blockade of autophagy enhances proapoptotic potential of BI-69A11, a novel Akt inhibitor, in colon carcinoma. Eur. J. Pharmacol. 2015 Aug 22 [PMID: 26306675] (WB, Human)
• Lee YH, Chen HY, Su LJ, Chueh PJ. Sirtuin 1 (SIRT1) Deacetylase Activity and NAD(+)/NADH Ratio Are Imperative for Capsaicin-Mediated Programmed Cell Death. J. Agric. Food Chem. 2015 Aug 26 [PMID: 26255724] (WB, Human)
  
  Details:
  ATG5 antibody was use for WB on lysates of MRC-5 normal human foetal lung fibroblast cells and adenocarcinomic human alveolar basal epithelial cell line A549 that were treated or not with Capsaicin or ethanol.
• Johansson I, Monsen VT, Pettersen K et al. The marine n-3 PUFA DHA evokes cytoprotection against oxidative stress and protein misfolding by inducing autophagy and NFE2L2 in human retinal pigment epithelial cells. Autophagy. 2015 Aug 03 [PMID: 26237736] (WB, Human)
  
  Details:
  ATG5 antibody used for WB on ARPE-19 cells that were transfected with ATG5 siRNA (Figure 4A).
• Dever SM, Rodriguez M, Lapierre J et al. Differing roles of autophagy in HIV-associated neurocognitive impairment and encephalitis with implications for morphine co-exposure. Front Microbiol 2015 [PMID: 26217309] (IHC)
• Pukab K, Goldbaum O, Richter-Landsberg C et al. Mitochondrial impairment and oxidative stress compromise autophagosomal degradation of alpha-synuclein in oligodendroglial cells J. Neurochem. 2015 Jul 24 [PMID: 26212128] (WB, Rat)
  
  Details:
  ATG5 antibody used for Western blot at a dilution of 1:500 on OLN-t40 cells (oligodendroglial cell line derived from rat brain glial cultures) that were transfected with Atg5 shRNA.
• Herpin A, Englberger E, Zehner M et al. Defective autophagy through epg5 mutation results in failure to reduce germ plasm and mitochondria. FASEB J 2015 Oct [PMID: 26183773]
• Yang S, Long LH, Li D et al. beta-Guanidinopropionic acid extends the lifespan of Drosophila melanogaster via an AMP-activated protein kinase-dependent increase in autophagy. Aging Cell 2015 Dec [PMID: 26120775] (WB)
• Till A, Saito R, Merkurjev D et al. Evolutionary trends and functional anatomy of the human expanded autophagy network. Autophagy 2015 [PMID: 26103419] (WB)
• Yao J, Jia L, Khan N et al. Deletion of autophagy inducer RB1CC1 results in degeneration of the retinal pigment epithelium Autophagy. 2015 Jun 03 [PMID: 26075877] (WB, Mouse)
• Xin W, Zhao X, Liu L et al. Acetyl-CoA carboxylase 2 suppression rescues human proximal tubular cells from palmitic acid induced lipotoxicity via autophagy. Biochem Biophys Res Commun 2015 Jul 31 [PMID: 26022126] (Human)
• Kim Y, Kang YS, Lee NY et al. Uvrag targeting by Mir125a and Mir351 modulates autophagy associated with Ewsr1 deficiency. Autophagy. 2015 May 04 [PMID: 25946189] (WB, Mouse)
• Wu Q, Jiang D, Huang C et al. Trehalose-mediated autophagy impairs the anti-viral function of human primary airway epithelial cells PLoS ONE 2015 Apr 17 [PMID: 25879848] (WB, Human)
• Gonnella R, Granato M, Farina A et al. PKC theta and p38 MAPK activate the EBV lytic cycle through autophagy induction. Biochim. Biophys. Acta. 2015 Mar 27 [PMID: 25827954] (WB, Human)
• Mohanty S, Jagannathan L, Ganguli G et al. A mycobacterial phosphoribosyltransferase promotes bacillary survival by inhibiting oxidative stress and autophagy pathways in macrophages and zebrafish J. Biol. Chem. 2015 Mar 30 [PMID: 25825498] (WB, Zebrafish)
• Sharifi MN, Mowers EE, Drake LE, Macleod KF. Measuring autophagy in stressed cells. Methods Mol. Biol. 2015 Mar 25 [PMID: 25804753] (WB, IHC-P, Human, Mouse)
• Roux C, Aligny C, Lesueur C et al. NMDA receptor blockade in the developing cortex induces autophagy-mediated death of immature cortical GABAergic interneurons: An ex vivo and in vivo study in Gad67-GFP mice. Exp. Neurol. 2015 Mar 17 [PMID: 25795167] (WB, Mouse)
  
  Details:
  ATG5 antibody used for WB on mouse cortical slices that were treated with MK801 (20 uM) alone or with 3-MA (30mM) for 6 hours.
• Prosser SL, Morrison CG. Centrin2 regulates CP110 removal in primary cilium formation. J. Cell Biol. 2015 Mar 16 [PMID: 25753040] (WB, Human)
• Carames B, Olmer M, Kiosses WB, Lotz M. The relationship of autophagy defects and cartilage damage during joint aging in a mouse model Arthritis & rheumatology. 2015 Feb 23 [PMID: 25708836] (IHC-P, Mouse)
• Reed M, Morris SH, Owczarczyk AB, Lukacs NW. Deficiency of autophagy protein Map1-LC3b mediates IL-17-dependent lung pathology during respiratory viral infection via ER stress-associated IL-1 Mucosal. Immunol 2015 Feb 11 [PMID: 25669150]
  
  Details:
  Using the DyLight 550 conjugated version of NB110-53818, catalog number NB110-53818R.
• Park E, Park MH, Na HS, Chung J. Xylitol induces cell death in lung cancer A549 cells by autophagy Biotechnol. Lett. 2015 Feb 04 [PMID: 25650339] (WB, Human)
  
  Details:
  ATG5 antibody used in WB application for the detection of Atg5- Atg12 conjugates in lysates of A549 cells which were subjected to induction of autophagy with Xylitol (cells pretreated with 5 mM of 3-MA for 1 hour prior to treatments with 1 uM rapamycin or 6% xylitol for 6 hours or 24 hours). Xylitol induced autophay to a level which was comparable to rapamycin and the data is shown in Figure 5c and 5d.
• Binotti B, Pavlos NJ, Riedel D et al. The GTPase Rab26 links synaptic vesicles to the autophagy pathway Elife. 2015 Feb 17 [PMID: 25643395] (WB, Human)
• Lin HH. In Vitro and in Vivo Atheroprotective Effects of Gossypetin against Endothelial Cell Injury by Induction of Autophagy Chem. Res. Toxicol. 2015 Jan 29 [PMID: 25622137] (WB, Human)
• Malhotra R, Warne JP, Salas E et al. Loss of Atg12, but not Atg5, in pro-opiomelanocortin neurons exacerbates diet-induced obesity Autophagy. 2015 Jan 13 [PMID: 25585051] (WB)
• Varga M, Fodor E, Vellai T. Autophagy in zebrafish. Methods. 2014 Dec 09 [PMID: 25498006] (WB, Zebrafish)
• Sun KT, Chen MY, Tu MG et al. MicroRNA-20a regulates autophagy related protein-ATG16L1 in hypoxia-induced osteoclast differentiation. Bone. 2014 Dec 05 [PMID: 25485521] (WB, Mouse)
  
  Details:
  ATG5 antibody used for WB on RAW264.7 cells subjected to RANKL and M-CSF mediated osteoclast differentiation as well as hypoxic stress (Fig. 1E )
• Kraya AA, Piao S, Xu X et al. Identification of secreted proteins that reflect autophagy dynamics within tumor cells. Autophagy 2015 [PMID: 25484078] (ELISA, Human)
• Kao C, Chao A, Tsai Cl et al. Bortezomib enhances cancer cell death by blocking the autophagic flux through stimulating ERK phosphorylation Cell Death Dis et al. 2014 Nov 07 [PMID: 25375375] (WB, Human)
  
  Details:
  ATG5 antibody used for WB on ovarian cancer TOV112D cells treated or not with bortezomib/BTZ.
• El-Hage N, Rodriguez M, Dever Sm et al. HIV-1 and morphine regulation of autophagy in microglia: Limited interactions in the context of HIV-1 infection and opioid abuse J. Virol. 2014 Oct 29 [PMID: 25355898] (WB, Human)
  
  Details:
  ATG5 antibody used for WB at 1:1000 dilution on lysates of primary human microglial cells that were treated or not with HIV-1SF162 macrophage (M)-tropic strain and/or Morphine for 1, 3 or 5 days (Figure 1B).
• Lin Hh, Lin Sm, Chung Y et al. Dynamic involvement of ATG5 in cellular stress responses. Cell Death Dis. 2014 Oct 24 [PMID: 25341032] (WB, IHC-P, Mouse)
• Rozman S, Yousefi S, Oberson K et al. The generation of neutrophils in the bone marrow is controlled by autophagy. Cell Death Differ. 2014 Oct 17 [PMID: 25323583] (WB, Mouse)
• Liao Cl, Chen Cm, Chang Yz et al. Pine (Pinus morrisonicola Hayata) needle Extracts Sensitize GBM8901 Human Glioblastoma Cells to Temozolomide by Downregulating Autophagy and O(6)-Methylguanine-DnA Methyltransferase Expression. J. Agric. Food Chem. 2014 Oct 29 [PMID: 25293350] (WB, Human)
• Granato M, Santarelli R, Farina A et al. EBV blocks the autophagic flux and appropriates the autophagic machinery to enhance viral replication. J Virol. 2014 Aug 20 [PMID: 25142602] (WB, Primate)
• Ligeon LA, Moreau K, Barois N et al. Role of VAMP3 and VAMP7 in the commitment of Yersinia pseudotuberculosis to LC3-associated pathways involving single- or double-membrane vacuoles. Autophagy 2014 Sep [PMID: 25046114] (WB)
• Montaigne D, Marechal X, Coisne A et al. Myocardial Contractile Dysfunction is Associated with Impaired Mitochondrial Function and Dynamics in Type 2 Diabetic but not in Obese Patients. Circulation. 2014 Jun 13 [PMID: 24928681] (WB, Human)
• Zielniok K, Motyl T, Gajewska M. Functional Interactions between 17beta-Estradiol and Progesterone Regulate Autophagy during Acini Formation by Bovine Mammary Epithelial Cells in 3D Cultures. BioMed Research International. 2014 May 07 [PMID: 24895572] (WB, Bovine)
• Yang Annan, Rajeshkumar N V, Wang Xiaoxu et al. Autophagy is critical for pancreatic tumor growth and progression in tumors with p53 alterations. Cancer Discov. 2014 May 29 [PMID: 24875860]
• Fang EF, Scheibye-Knudsen M, Brace LE et al. Defective Mitophagy in XPA via PARP-1 Hyperactivation and NAD(+)/SIRT1 Reduction. Cell. 2014 May 08 [PMID: 24813611] (WB, Mouse)
• Bejarano E, Yuste A, Patel B et al. Connexins modulate autophagosome biogenesis. Nat. Cell Biol. 2014 Apr 06 [PMID: 24705551] (WB, IP, Mouse)
• Jabir MS, Ritchie ND, Li D et al. Caspase-1 Cleavage of the TLR Adaptor TRIF Inhibits Autophagy and beta-Interferon Production during Pseudomonas aeruginosa Infection. Cell Host Microbe [PMID: 24528867] (WB, Mouse)
• Haldar AK, Piro AS, Pilla DM, Coers J. The E2-like conjugation enzyme Atg3 promotes binding of IRG and Gbp proteins to Chlamydia- and Toxoplasma-containing vacuoles and host resistance. PLoS ONE 2014 Jan 27 [PMID: 24466199] (WB, Mouse)
• Feng W, Chang C, Luo D et al. Dissection of autophagy in human platelets. Autophagy 2014 Apr [PMID: 24458007] (WB)
• Choi Soyoung, Shin Hyejin, Song Haengseok, Lim Hyunjung Jade. Suppression of autophagic activation in the mouse uterus by estrogen and progesterone. J Endocrinol. 2014 Mar 07 [PMID: 24443716] (Mouse)
• Lee E, Koo Y, Ng A et al. Autophagy is essential for cardiac morphogenesis during vertebrate development. Autophagy 2014 Apr [PMID: 24441423] (WB)
• Townsend KN, Spowart JE, Huwait H et al. Markers of T Cell Infiltration and Function Associate with Favorable Outcome in Vascularized High-Grade Serous Ovarian Carcinoma. PLoS One 2013 Dec 23 [PMID: 24376535] (WB, Mouse)
• Varga M, Sass M, Papp D et al. Autophagy is required for zebrafish caudal fin regeneration. Cell Death Differ 2013 Dec 6 [PMID: 24317199] (WB, Zebrafish)
• Liu Y, Shoji-Kawata S, Sumpter RM Jr et al. Autosis is a Na+,K+-ATPase-regulated form of cell death triggered by autophagy-inducing peptides, starvation, and hypoxia-ischemia. Proc Natl Acad Sci U S A. 2013 Nov 25 [PMID: 24277826] (WB, Mouse)
• Pei J, Zhao M, Ye Z et al. Autophagy enhances the replication of classical swine fever virus in vitro. Autophagy 2014 Jan [PMID: 24262968] (WB)
• Martinez-Lopez N, Athonvarangkul D, Mishall P et al. Autophagy proteins regulate ERK phosphorylation. Nat Commun. 2013 Nov 18 [PMID: 24240988] (ICC/IF, WB, Mouse)
• Cha HH, Hwang JR, Kim HY et al. Autophagy Induced by Tumor Necrosis Factor alpha Mediates Intrinsic Apoptosis in Trophoblastic Cells. Reprod Sci. 2013 Nov 6 [PMID: 24198074] (WB, Human)
• Hollomon MG, Gordon N, Santiago-O Farrill JM, Kleinerman ES. Knockdown of autophagy-related protein 5, ATG5, decreases oxidative stress and has an opposing effect on camptothecin-induced cytotoxicity in osteosarcoma cells. BMC Cancer. 2013 Oct 26 [PMID: 24160177] (WB, Mouse)
• Roy K, Raychaudhuri M, Chakrabarti O, Mukhopadhyay D. Growth Factor Receptor-Bound Protein 2 Promotes Autophagic Removal of Amyloid-beta Protein Precursor Intracellular Domain Overload in Neuronal Cells. J Alzheimers Dis. 2013 Oct 7 [PMID: 24100123]
• Granato M, Santarelli R, Lotti LV et al. JNK and Macroautophagy Activation by Bortezomib Has a Pro-Survival Effect in Primary Effusion Lymphoma Cells. PLoS One. 2013 Sep 26 [PMID: 24086672] (WB, Human)
• Lucin Kurt M, O'brien Caitlin E, Bieri Gregor et al. Microglial beclin 1 regulates retromer trafficking and phagocytosis and is impaired in Alzheimer's disease. Neuron 2013 [PMID: 24012002] (IHC, Mouse)
• Cufi S, Vazquez-Martin A, Oliveras-Ferraros C et al. The anti-malarial chloroquine overcomes Primary resistance and restores sensitivity to Trastuzumab in HER2-positive breast cancer. Sci Rep. 2013 Aug 22 [PMID: 23965851] (WB, Human)
• Artal-Martinez de Narvajas A, Gomez TS, Zhang JS et al. Epigenetic regulation of autophagy by the methyltransferase G9a. Mol Cell Biol 2013 Oct [PMID: 23918802]
• Martinez-Lopez N, Athonvarangkul D, Sahu S et al. Autophagy in Myf5+ progenitors regulates energy and glucose homeostasis through control of brown fat and skeletal muscle development. EMBO Rep 2013 Aug 2 [PMID: 23907538] (WB, Mouse)
• Essick EE, Wilson RM, Pimentel DR et al. Adiponectin Modulates Oxidative Stress-Induced Autophagy in Cardiomyocytes. PLoS One 2013 Jul 19 [PMID: 23894332] (WB, Rat)
• Reed M, Morris SH, Jang S et al. Autophagy-Inducing Protein Beclin-1 in Dendritic Cells Regulates CD4 T Cell Responses and Disease Severity during Respiratory Syncytial Virus Infection. J Immunol 2013 Jul 26 [PMID: 23894198] (ICC/IF, Mouse)
  
  Details:
  Using the DyLight 550 conjugated version of NB110-53818, catalog number NB110-53818R.
• Morgan-Bathke M, Lin HH, Chibly AM et al. Deletion of ATG5 Shows a Role of Autophagy in Salivary Homeostatic Control. J Dent Res 2013 Jul 24 [PMID: 23884556] (WB, Mouse)
• Zhang C, He Y, Okutsu M et al. Autophagy is involved in adipogenic differentiation by repressesing proteasome-dependent PPARy2 degradation. Am J Physiol Endocrinol Metab. 2013 Aug [PMID: 23800883] (WB, Mouse)
• Benjamin JL, Sumpter R Jr, Levine B, Hooper LV. Intestinal Epithelial Autophagy Is Essential for Host Defense against Invasive Bacteria. Cell Host Microbe 2013 Jun 12 [PMID: 23768496] (WB, Mouse)
• Ciccia F, Accardo-Palumbo A, Rizzo A et al. Evidence that autophagy, but not the unfolded protein response, regulates the expression of IL-23 in the gut of patients with ankylosing spondylitis and subclinical gut inflammation. Ann Rheum Dis 2013 Jun 5 [PMID: 23740229]
• Sorrentino V, Nelson JK, Maspero E et al. The LXR-IDOL axis defines a clathrin, caveolae, and dynamin-independent endocytic route for LDLR internalization and lysosomal degradation. J Lipid Res 2013 Jun 3 [PMID: 23733886] (WB, Human)
• Fedorova LV, Sodhi K, Gatto-Weis C et al. Peroxisome Proliferator-Activated Receptor delta Agonist, HPP593, Prevents Renal Necrosis under Chronic Ischemia. PLoS One 2013 May 15 [PMID: 23691217] (WB, Rat)
• Notte A, Ninane N, Arnould T, Michiels C. Hypoxia counteracts taxol-induced apoptosis in MDA-MB-231 breast cancer cells: role of autophagy and JNK activation. Cell Death Dis 2013 May 16 [PMID: 23681233] (WB, Human)
• Laforge M, Limou S, Harper F et al. DRAM Triggers Lysosomal Membrane Permeabilization and Cell Death in CD4(+) T Cells Infected with HIV. PLoS Pathog 2013 May [PMID: 23658518] (WB, Human)
• Amir M, Zhao E, Fontana L et al. Inhibition of hepatocyte autophagy increases tumor necrosis factor-dependent liver injury by promoting caspase-8 activation Cell Death Differ 2013 Mar 22 [PMID: 23519075] (WB, Mouse)
• Long L, Yang X, Southwood M et al. Chloroquine Prevents Progression of Experimental Pulmonary Hypertension via Inhibition of Autophagy and Lysosomal Bmpr-II Degradation. Circ Res 2013 Feb 27 [PMID: 23446737] (WB, Rat)
• Yen YT, Yang HR, Lo HC et al. Enhancing autophagy with activated protein C and rapamycin protects against sepsis-induced acute lung injury. Surgery 2013 Feb 19 [PMID: 23434181] (WB, IHC-P, Mouse)
• Fu M, St-Pierre P, Shankar J et al. Regulation of mitophagy by the Gp78 E3 ubiquitin ligase. Mol Biol Cell 2013 Apr [PMID: 23427266] (WB, Human)
• Weber K, Schilling JD. Distinct Lysosome Phenotypes Influence Inflammatory Function in Peritoneal and Bone Marrow-Derived Macrophages. International Journal of Inflammation. 2014 Dec 23 (WB, Mouse)
  
  Details:
  ATG5 antibody used for WB on the total cellular protein lysates of Peritoneal macrophage / pMACs or Bone marrow-derived macrophage / BMDMs isolated from C57BL/6 mice (Figure 4g)
• Shoji-Kawata S, Sumpter R, Leveno M et al. Identification of a candidate therapeutic autophagy-inducing peptide. Nature. 2013 Feb 14 [PMID: 23364696]
• Brennan GP, Jimenez-Mateos EM, McKiernan RC et al. Transgenic Overexpression of 14-3-3 Zeta Protects Hippocampus against Endoplasmic Reticulum Stress and Status Epilepticus In Vivo. PLoS One 2013 [PMID: 23359526] (WB, Mouse)
• Shi JM, Bai LL, Zhang DM et al. Saxifragifolin D induces the interplay between apoptosis and autophagy in breast cancer cells through ROS-dependent endoplasmic reticulum stress. Biochem Pharmacol 2013 Jan 21 [PMID: 23348250] (WB, Human)
• Chen Y, Sawada O, Kohno H et al. Autophagy protects the retina from light-induced degeneration. J Biol Chem 2013 Jan 22 [PMID: 23341467] (WB, Mouse)
• Zhu S, Cao L, Yu Y et al. Inhibiting autophagy potentiates the anticancer activity of IFN1@/ IFNalpha in chronic myeloid leukemia cells. Autophagy 2012 Dec 14 [PMID: 23242206] (WB, Human)
• Kang R, Tang D, Lotze MT, Zeh Iii HJ. Autophagy is required for IL-2-mediated fibroblast growth Exp Cell Res 2012 Nov 27 [PMID: 23195496] (Mouse)
• Curtis S, Jones CJ, Garrod A et al. Identification of Autophagic Vacuoles and Regulators of Autophagy in Villous Trophoblast from Normal Term Pregnancies and in Fetal Growth Restriction J Matern Fetal Neonatal Med 2012 Oct 8 [PMID: 23039021] (IHC, Human)
• Fujii S, Hara H, Araya J et al. Insufficient autophagy promotes bronchial epithelial cell senescence in chronic obstructive pulmonary disease Oncoimmunology 2012 Aug 1 [PMID: 22934255] (WB, Human)
• Zang Y, Thomas SM, Chan ET et al. Carfilzomib and ONX 0912 Inhibit Cell Survival and Tumor Growth of Head and Neck Cancer and Their Activities Are Enhanced by Suppression of Mcl-1 or Autophagy Clin Cancer Res 2012 Oct 15 [PMID: 22929803] (WB, Human)
• Chakrabarti A, Ghosh PK, Banerjee S et al. RNase L Triggers Autophagy in Response to Viral Infections J Virol 2012 Oct [PMID: 22875977] (WB, Mouse)
• Cannizzo ES, Clement CC, Morozova K et al. Age-Related Oxidative Stress Compromises Endosomal Proteostasis Cell Rep 2012 Jul 26 [PMID: 22840404] (WB, Mouse)
• Sandilands E, Serrels B, Wilkinson S, Frame MC. Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells EMBO Rep 2012 Jun 26 [PMID: 22732841] (WB, Mouse)
• Garcia AG, Wilson RM, Heo J et al. Interferon-gamma ablation exacerbates myocardial hypertrophy in diastolic heart failure Am J Physiol Heart Circ Physiol 2012 Sep [PMID: 22730392] (WB, Mouse)
• Kang R, Loux T, Tang D, Schapiro NE, Vernon P, Livesey KM, Krasinskas A, Lotze MT, Zeh HJ 3rd. The expression of the receptor for advanced glycation endproducts (RAGE) is permissive for early pancreatic neoplasia. Proc Natl Acad Sci U S A;109(18):7031-6. 2012 May 1. [PMID: 22509024] (WB, Mouse)
• Razani B, Feng C, Coleman T et al. Autophagy Links Inflammasomes to Atherosclerotic Progression Cell Metabolism 2012 Apr 4 [PMID: 22440612] (WB, Mouse)
• Sharara-Chami RI, Zhou Y, Ebert S et al. Epinephrine deficiency results in intact glucose counter-regulation, severe hepatic steatosis and possible defective autophagy in fasting mice The International Journal of Biochemistry & Cell Biology 2012 Jun [PMID: 22405854] (WB, Mouse)
• Yu Y, Yang L, Zhao M et al. Targeting microRNA-30a-mediated autophagy enhances imatinib activity against human chronic myeloid leukemia cells Leukemia 2012 Mar 7 [PMID: 22395361] (WB, Human)
• Zhang Y, Morgan MJ, Chen K et al. Induction of autophagy is essential for monocyte-macrophage differentiation. Blood. 2012 Jan 5. [PMID: 22223827]
• Vigano A, Vasso M, Caretti A, Bravata V, Terraneo L, Fania C, Capitanio D, Samaja M, Gelfi C. Protein modulation in mouse heart under acute and chronic hypoxia. Proteomics;11(21):4202-17. 2011 Nov. [PMID: 22150682] (WB, Human)
• Cufi S, Vazquez-Martin A, Oliveras-Ferraros C et al. Autophagy positively regulates the CD44(+) CD24(-/low) breast cancer stem-like phenotype. Cell Cycle 2011 Nov 15 [PMID: 22127234] (Human)
• Lee H, Noh J-Y, Oh Y et al. IRE1 plays an essential role in ER stress-mediated aggregation of mutant huntingtin via the inhibition of autophagy flux. Human Molecular Genetics. 2011 Sep 27. [PMID: 21954231]
• Li JK, Liang JJ, Liao CL, Lin YL. Autophagy is involved in the early step of Japanese encephalitis virus infection. Microbes Infect;14(2):159-68. 2012 Feb. [PMID: 21946213]
• Ling Y-H, Aracil M, Zou Y et al. PM02734 Induces Caspase-Independent Cell Death Associated with Features of Autophagy, Inhibition of the Akt/mTOR Signaling Pathway, and Activation of DAP Kinase. Clin Cancer Res;17(16):5353-66. 2011 Aug 15. [PMID: 21690574]
• Raveh B, London N, Zimmerman L, Schueler-Furman O. Rosetta FlexPepDock ab-initio: simultaneous folding, docking and refinement of peptides onto their receptors. PLoS One 2011 [PMID: 21572516] (WB)
• Dash R, Azab B, Quinn B et al. Apogossypol derivative BI-97C1 (Sabutoclax) targeting Mcl-1 sensitizes prostate cancer cells to mda-7/IL-24-mediated toxicity Proc Natl Acad Sci U S A 2011 May 24 [PMID: 21555592] (WB, Human)
• Lee J-E, Oh H-A, Song H et al. Autophagy Regulates Embryonic Survival During Delayed Implantation. Endocrinology;152(5):2067-75. 2011 May. [PMID: 21363932] (ICC/IF, Mouse)
• Elgendy M, Sheridan C, Brumatti G, Martin SJ. Oncogenic Ras-Induced Expression of Noxa and Beclin-1 Promotes Autophagic Cell Death and Limits Clonogenic Survival. Mol Cell. 2011 Feb 23. [PMID: 21353614]
• Herd HL, Malugin A, Ghandehari H. Silica nanoconstruct cellular toleration threshold in vitro. J Control Release;153(1):40-8. 2011 Jul 15. [PMID: 21342660] (WB, Human, Mouse)
• Wang RC, Levine B. Calcipotriol Induces Autophagy in HeLa Cells and Keratinocytes. J Invest Dermatol. 2011 [PMID: 21228817]
• Mostowy S, Bonazzi M, Hamon MA et al. Entrapment of intracytosolic bacteria by septin cage-like structures. Cell Host Microbe 2010 Nov 18 [PMID: 21075354]
• Brady OA, Meng P, Zheng Y et al. egulation of TDP-43 aggregation by phosphorylation and p62/SQSTM1 J Neurochem 2011 Jan [PMID: 21062285] (WB, ICC/IF, Mouse, Primate)
• Travassos LH, Carneiro LA, Ramjeet M et al. Nod1 and Nod2 direct autophagy by recruiting ATG16L1 to the plasma membrane at the site of bacterial entry. Nat Immunol 2010 Jan [PMID: 19898471]
• Fujimoto K, Hanson PT, Tran H et al. Autophagy Regulates Pancreatic Beta Cell Death in Response to Pdx1 Deficiency and Nutrient Deprivation. J Biol Chem;284(40):27664-27673. 2009 [PMID: 19654319]
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• English L, Chemali M, Duron J et al. Autophagy enhances the presentation of endogenous viral antigens on MHC class I molecules during HSV-1 infection. Nat Immunol 2009 May [PMID: 19305394] (WB, Mouse)
• Cadwell K, Liu JY, Brown SL et al. A key role for autophagy and the autophagy gene Atg16l1 in mouse and human intestinal Paneth cells. Nature;456(7219):259-63. 2008 Nov 13. [PMID: 18849966]
• Pyo, J, Nah, J, Kim, H et al. Compensatory activation of ERK1/2 in Atg5-deficient mouse embryo fibroblasts suppresses oxidative stress-induced cell death. Autophagy;4(3):315-21. 2008 Apr 1. [PMID: 18196969] (WB, Mouse, Human)
• Wang, Y, R Singh et al. Loss of macroautophagy promotes or prevents fibroblast apoptosis depending on the death stimulus. J Biol Chem;283 (8):4766-77. 2008 Feb 22. [PMID: 18073215]

Reviews for ATG5 Antibody (NB110-53818) (6)

Average Rating: 4.2

(Based on 6 reviews)

We have 6 reviews tested in 3 species: Human, Mouse, Rat.

reviewed by: Anonymous WB Mouse 03/31/2018

Summary

• Application: Western Blot
• Sample Tested: Mouse Colon
• Species: Mouse
• Lot: M-1

reviewed by: Anonymous WB Human 12/15/2017

Summary

• Application: Western Blot
• Sample Tested: human parimary hepatocytes,Human primary hepatocytes
• Species: Human
• Lot: K-3

Comments

• Comments: 100 ng/ml LPS treated human primary hepatocytes

reviewed by: Merissa Olmer IHC-P 10/04/2014

Summary

• Application: Immunohistochemistry-Paraffin
• Lot: E-3
• Comments: These tissue samples were fixed with zinc buffered formalin.

reviewed by: Ann Marie Santiago WB Mouse 09/21/2012

Summary

• Application: Western Blot
• Sample Tested: Murine hepatoma
• Species: Mouse
• Lot: e-2
• Comments: May be used at lower dilution. Overnight incubation @ 4deg C is also adequate.

Blocking

• Blocking Details: Milk, 5% in TBS-Tween

Primary Anitbody

• Dilution Ratio: 1:500 in TBS-Tween, 1hr @RT

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG alkaline phosphatase linked
• Secondary Manufacturer Cat#: NIF1317
• Secondary Concentration: 1:10000

Details

• Detection Notes: GE/Amersham ECF Detection, Molecular Dynamics Storm 840 scanner

Comments

• Comments: May be used at lower dilution. Overnight incubation @ 4deg C is also adequate.

reviewed by: Anonymous WB Rat 12/20/2011

Summary

• Application: Western Blot
• Sample Tested: Rat Cardiomyocyte
• Species: Rat
• Comments: The major band was observed, but sometimes there was a non-specific band detected around 4 KDa

Blocking

• Blocking Details: Blocking Buffer: 5% Milk in TBST, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 C

Secondary Antibody

• Secondary Description: Secondary Ab: Goat anti-Rabbit, Secondary Ab Dilution Ratio: 1:5000

Details

• Detection Notes: Detection Method: ECL

Comments

• Comments: The major band was observed, but sometimes there was a non-specific band detected around 4 KDa

reviewed by: Anonymous WB Mouse 11/16/2011

Summary

• Application: Western Blot
• Sample Tested: Mouse
• Species: Mouse

Blocking

• Blocking Details: Blocking Buffer: BSA

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: anti-rabbit HRP

Details

• Detection Notes: Detection Method: ECL

FAQs for ATG5 Antibody (NB110-53818). (Showing 1 - 5 of 5 FAQs).

1. I am interested in testing your anti ATG5 (NB110-53818SS) for Immunofluorescence in MDCK cell (canine epithelial cell line). I noticed you report reactivity of the antibody against several species but not dog. Do you know if the Ab will react against canine genome? Also for IF what fixation do you recommend?
   • The applications and species listed for any one of our antibodies are ones we have tested and validated. Canine is not listed as it has not been tested and therefore we cannot guarantee its cross reactivity. If you would be interested in testing this antibody in Immunofluorescence, we can recommend our Innovator's Reward. Our Innovator's Reward is offered to reward researchers for testing new species and applications with our products. Learn more about our Innovators Reward Program. Also, please use this link to view our Immunofluorescence protocol which explains the fixation method that we recommend.
2. Our customer is interested in this ATG5 antibody (cat. no. NB110-53818). Would you please confirm that this antibody would recognize ATG5 alone, as well as ATG5-ATG12 conjugate?
   • The antibody was raised against a peptide corresponding to the N-terminal of ATG5. It should recognize ATG5 alone but it appears that ATG5 is rarely found alone in the cell.
3. Could you help me with the following question about ATG5 antibody (Cat. NB110-53818) that I bought from you? The arrow in the data sheet points to Atg5/Atg12 band of 56kDa, there is also a band of 33kDa which I guess is the Atg5? Could you excuse my ignorance and explain what is the meaning of detecting the Atg5/Atg12 conjugate in preference to Atg5, I'm obviously missing something and Wikipedia has nothing on the subject. Do I guess correctly that the antibody does not cross-react with ATG12?
   • The western blot image you are referring to illustrates the Atg5/Atg12 conjugate, which is formed during autophagy. During autophagy Atg5 is covalently bonded with Atg12, this is a ubiquitous reaction. The lysate that was used was meant to illustrate the ability to detect autophagy by WB via Atg5/Atg12. It is possible to detect free floating Atg5 in cell lysates although it is not seen in this Western blot. The lower 2 bands do not correspond to Atg5. Our Atg5 antibody is not able to detect Atg12, which free floating runs at about 23 kDa.
4. As you mention in 3rd FAQs (Frequently asked questions) section, that this antibody can detect free ATG5 in cell lysates (although there is no western blot image). Does this antibody can detect free ATG5 monomer in Hela or HEK 293 cells?
   • With regard to our product NB110-53818 and the FAQ #3, this antibody is able to detect the ATG5 (unconjugated form) ~33kDa and the conjugated form ~56kDa. In the western blot image we have on our website, we show the antibody's ability to detect the autophagy induced conjugate ATG5/ATG12. We also have in a customer review, an image of both the conjugate and unconjugated ATG5 in rat Cardiomyocyte. We also have quite a few publications that show this antibody working in western blot with mouse and human samples. I am not sure if any of the publications have specifically tested HeLa or HEK 293 cells, but if the ATG5 protein is present, this product will be able to detect it in western blot.
5. Would you please help confirm if NB110-53818 has been tested by IHC with frozen sections?
   • NB110-53818 has been tested with paraffin-embedded tissue, but we have not validated this antibody for IHC with frozen samples.

Bioinformatics

• Gene Symbol: ATG5