ATG5 Antibody

Images

 
Genetic Strategies: 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 ...read more
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 ...read more
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 ...read more
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 ...read more
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 ...read more

Product Details

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

Genetic Strategies

   

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
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

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)
Reviewed Applications
Read 6 Reviews rated 4.2
using
NB110-53818 in the following applications:

Publications
Read Publications using
NB110-53818 in the following applications:

  • 1 publication
  • EM
    1 publication
  • 1 publication
  • 7 publications
  • IHC
    4 publications
  • 6 publications
  • IP
    1 publication
  • PLA
    1 publication
  • RIA
    1 publication
  • WB
    108 publications

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.

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NBP1-88883
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NBP1-55202
Species: Hu
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NB110-53818
Species: Hu, Mu, Rt, Po, Bv, Pm, Xp, Ze
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP, PLA, RIA, KO

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

We have publications tested in 6 confirmed species: Human, Mouse, Rat, Bovine, Primate, Zebrafish.

We have publications tested in 10 applications: ELISA, EM, Flow, ICC/IF, IHC, IHC-P, IP, PLA, RIA, WB.


Filter By Application
ELISA
(1)
EM
(1)
Flow
(1)
ICC/IF
(7)
IHC
(4)
IHC-P
(6)
IP
(1)
PLA
(1)
RIA
(1)
WB
(108)
All Applications
Filter By Species
Human
(42)
Mouse
(60)
Rat
(6)
Bovine
(4)
Primate
(2)
Zebrafish
(7)
All Species
Showing Publications 1 - 10 of 157. Show All 157 Publications.
Publications using NB110-53818 Applications Species
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) WB Mouse
Gibson J. In vivo imaging and analysis of host-pathogen interactions of intracellular pathogens Thesis 2017 (WB, Zebrafish) 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) 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) 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) 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) 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) 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) 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]
Show All 157 Publications.

Reviews for ATG5 Antibody (NB110-53818) (6) 4.26

Average Rating: 4.2
(Based on 6 reviews)
We have 6 reviews tested in 3 species: Human, Mouse, Rat.

Reviews using NB110-53818:
Filter by Applications
WB
(5)
IHC-P
(1)
All Applications
Filter by Species
Human
(1)
Mouse
(3)
Rat
(1)
All Species
Images Ratings Applications Species Date Details
 
reviewed by:
Anonymous
WB Mouse 03/31/2018
View

Summary

ApplicationWestern Blot
Sample TestedMouse Colon
SpeciesMouse
LotM-1
Western Blot ATG5 NB110-53818
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reviewed by:
Anonymous
WB Human 12/15/2017
View

Summary

ApplicationWestern Blot
Sample Testedhuman parimary hepatocytes,Human primary hepatocytes
SpeciesHuman
LotK-3

Comments

Comments100 ng/ml LPS treated human primary hepatocytes
Immunohistochemistry-Paraffin ATG5 NB110-53818
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reviewed by:
Merissa Olmer
IHC-P 10/04/2014
View

Summary

ApplicationImmunohistochemistry-Paraffin
LotE-3
CommentsThese tissue samples were fixed with zinc buffered formalin.
Western Blot ATG5 NB110-53818
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reviewed by:
Ann Marie Santiago
WB Mouse 09/21/2012
View

Summary

ApplicationWestern Blot
Sample TestedMurine hepatoma
SpeciesMouse
Lote-2
CommentsMay be used at lower dilution. Overnight incubation @ 4deg C is also adequate.

Blocking

Blocking DetailsMilk, 5% in TBS-Tween

Primary Anitbody

Dilution Ratio1:500 in TBS-Tween, 1hr @RT

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG alkaline phosphatase linked
Secondary Manufacturer Cat#NIF1317
Secondary Concentration1:10000

Details

Detection NotesGE/Amersham ECF Detection, Molecular Dynamics Storm 840 scanner

Comments

CommentsMay be used at lower dilution. Overnight incubation @ 4deg C is also adequate.
Western Blot ATG5 NB110-53818
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reviewed by:
Anonymous
WB Rat 12/20/2011
View

Summary

ApplicationWestern Blot
Sample TestedRat Cardiomyocyte
SpeciesRat
CommentsThe major band was observed, but sometimes there was a non-specific band detected around 4 KDa

Blocking

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

Primary Anitbody

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

Secondary Antibody

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

Details

Detection NotesDetection Method: ECL

Comments

CommentsThe major band was observed, but sometimes there was a non-specific band detected around 4 KDa
 
reviewed by:
Anonymous
WB Mouse 11/16/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse

Blocking

Blocking DetailsBlocking Buffer: BSA

Primary Anitbody

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

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-rabbit HRP

Details

Detection NotesDetection Method: ECL

Product General Protocols

View specific protocols for ATG5 Antibody (NB110-53818): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

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.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB110-53818AF405
Alexa Fluor 488 Labeled NB110-53818AF488
Alexa Fluor 647 Labeled NB110-53818AF647
Alexa Fluor 700 Labeled NB110-53818AF700
Biotin Labeled NB110-53818B
DyLight 405 Labeled NB110-53818V
DyLight 488 Labeled NB110-53818G
DyLight 550 Labeled NB110-53818R
DyLight 650 Labeled NB110-53818C
FITC Labeled NB110-53818F
HRP Labeled NB110-53818H

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Blogs on ATG5. Showing 1-10 of 26 blog posts - Show all blog posts.

Losing memory: Toxicity from mutant APP and amyloid beta explain the hippocampal neuronal damage in Alzheimer's disease
By Jamshed Arslan Pharm.D. Alzheimer's disease (AD) is an irreversible brain disorder that destroys memory and thinking skills. The telltale signs of AD brains are extracellular deposits of amyloid beta (a polypepti...  Read full blog post.


  Read full blog post.

Autophagy independent roles of the core ATG proteins
By Christina Towers, PhD. Autophagy and ATG ProteinsAutophagy is a nutrient recycling process that cells use to fuel metabolism, particularly in response to nutrient deprivation.  It is critical for removal of dam...  Read full blog post.

Key Targets in Apoptosis, Necroptosis, and Autophagy
Cell death/recycling pathways such as apoptosis, necroptosis, and autophagy are an integral part of the growth, development, homeostasis as well as the pathophysiology in the life of living organisms. These signaling pathways are highly regulated and...  Read full blog post.

The role of Parkin and autophagy in retinal pigment epithelial cell (RPE) degradation
The root of Parkinson’s disease (PD) points to a poorly regulated electron transport chain leading to mitochondrial damage, where many proteins need to work cohesively to ensure proper function.  The two key players of this pathway are PINK1,...  Read full blog post.

The use of actin as a loading control in research on fruiting-body development and vegetative growth in Sordaria macrospora research
Sordaria macrospora is a filamentous fungus that serves as very useful system for scientific research due to a short life cycle and easy manipulation.  Just like any other model organism, it is important to have an effective loading control to va...  Read full blog post.

Required proteins for p62/SQSTM1 regulation and a role for p62/SQSTM1 in neuronal autophagy
Autophagy is a crucial cellular process that clears the cell of protein aggregates, toxins, and damaged cell products. Accumulation of toxins, damaged cell products and unwanted proteins has been proven to play a role in aging and many forms of dis...  Read full blog post.

The role of LC3B and autophagy in alcohol induced liver disease
Autophagy is a crucial intracellular pathway that manages the degradation and recycling of long-lived proteins in the cell. The LC3 (or light chain 3) family is composed of three members, LC3A, LC3B and LC3C. Upon autophagy induction, LC3 is cleave...  Read full blog post.

WIPI1 - An essential regulator of early autophagosome assembly
WD repeat domain phosphoinositide-interacting protein 1 (WIPI) is involved in the lysosomal degradation of cytoplasmic components during starvation-induced autophagy. WIPI1 is a seven bladed beta-propeller protein that provides a scaffold for the ...  Read full blog post.

ATG4C - A regulator of the early steps of autophagosome assembly
Autophagy is an important cellular process that maintains homeostasis by degrading and recycling damaged proteins and organelles. Autophagy receptors, such as p62/SQSTM1, recognize these intracellular cargo and mediate their engulfment by the doubl...  Read full blog post.

Showing 1-10 of 26 blog posts - Show all blog posts.

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Recent Reviews

4.2
5
1
4
5
3
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2
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Anonymous
03/31/2018
Application: WB
Species: Mouse

Anonymous
12/15/2017
Application: WB
Species: Human

Merissa Olmer
10/04/2014
Application: IHC-P
Species:

Bioinformatics

Gene Symbol ATG5