ATG5 Antibody

Images

 
Genetic Strategies: Western Blot: ATG5 Antibody [NB110-53818] - Western blot analysis in mouse wildtype ES cell lysate (Lane 1) using NB110-53818. Lane 2 is a mouse ATG5 KO ES cell lysate (negative control). ...read more
Genetic Strategies: Western Blot: ATG5 Antibody [NB110-53818] - ERK phosphorylation in Atg5-/-cells depends on nutrient availability. Serum-deprived Atg5-/- MEFs display decreased ERK and p90RSK phosphorylation. ...read more
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - ICC/IF analysis of SY5Y cells at 1:250. Incubated overnight at 4 degrees. Photo courtesy of an anonymous collaborator.
Immunohistochemistry: ATG5 Antibody [NB110-53818] - Immunohistochemistry analysis of human liver hepatocytes at 2.5 ug/mL. 40X magnification.
Immunocytochemistry/ Immunofluorescence: ATG5 Antibody [NB110-53818] - ERK2 utilizes kinase-docking domains to interact with ATG5-ATG12 and LC3-II.(a) Mutations in FRS on ERK2 decrease colocalization of ERK2 with ...read more
Western Blot: ATG5 Antibody [NB110-53818] - Western blot analysis of total protein from Human HeLa and A431 and Mouse MEF cells. Lysates were separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked ...read more
Biological Strategies: Western Blot: ATG5 Antibody [NB110-53818] - IFN-gamma increased rates of autophagy in primary BMECs in vitro. BMECs were treated with IFN-gamma at 2.5, 5, 10 or 20 ng/mL. At the end of the ...read more
Immunohistochemistry: ATG5 Antibody [NB110-53818] - Immunohistochemistry analysis of mouse intestine using DAB with hematoxylin counterstain.
Immunohistochemistry-Paraffin: ATG5 Antibody [NB110-53818] - Immunohistochemistry analysis of mouse knee. IHC-P image submitted by a 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
Genetic Strategies: Knockdown Validated: ATG5 Antibody [NB110-53818] - Knockdown of autophagy-related gene 5 (ATG5) inhibits trehalose-induced autophagy in normal human primary airway epithelial cells. Normal ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Fi, Gp, Pm, Xp, ZeSpecies Glossary
Applications WB, Simple Western, ELISA, EM, Flow, IB, ICC/IF, IHC, IHC-P, IP, PLA, RIA, KD, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated

ATG5 Antibody Summary

Immunogen
This ATG5 Antibody was made to a synthetic peptide of 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
  • Electron Microscopy
  • ELISA
  • Flow Cytometry
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:250
  • Immunohistochemistry 1:400
  • Immunohistochemistry-Paraffin 1:400
  • Immunoprecipitation 1:10 - 1:500
  • Proximity Ligation Assay
  • Radioimmunoassay
  • Knockdown Validated
  • 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). Use in immunoblotting reported in scientific literature (PMID: 23918802). Use in ELISA reported in scientific literature (PMID: 25484078). Use in electron microscopy reported in scientific literature (PMID: 27219062). Use in ICC/IF was reported in scientific literature (PMID: 18073215).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Theoretical MW
32 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
ATG5 Overexpression Lysate
Control Peptide
ATG5 Protein (NB110-53818PEP)
Reviewed Applications
Read 7 Reviews rated 4.1
using
NB110-53818 in the following applications:

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

  • 1 publication
  • EM
    1 publication
  • 1 publication
  • 13 publications
  • IHC
    8 publications
  • 9 publications
  • IP
    2 publications
  • KO
    1 publication
  • PLA
    1 publication
  • RIA
    1 publication
  • WB
    145 publications

Reactivity Notes

Fish reactivity reported in scientific literature (PMID: 26183773). Guinea Pig reactivity reported in scientific literature (PMID: 30766882).

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, S. Cerevisiae)-Like
  • APG5 autophagy 5-like (S. cerevisiae)
  • APG5
  • APG5L
  • APG5-LIKE
  • ASP
  • ASPAPG5-LIKE
  • ATG5 autophagy related 5 homolog (S. cerevisiae)
  • ATG5 Autophagy Related 5 Homolog
  • ATG5
  • Autophagy protein 5
  • Autophagy Related 5
  • HAPG5
  • SCAR25

Background

Atg5, Autophagy related 5 or autophagy protein 5 (theoretical molecular weight 32 kDa), belongs to a group of core autophagy-related proteins first identified in yeast and later in eukaryotic cells. Atg proteins play essential roles in the process or macroautophagy. Atg5 is considered a core autophagy protein, for its role in the formation of the autophagosome, a double membrane vesicle which engulfs proteins and organelles for delivery to the lysosome and subsequent degradation (1). Atg5 participates in the process of phagophore elongation by interacting with the ubiquitin-like protein Atg12. Formation of the Atg12-Atg5 conjugate is dependent on the activities of Atg7 (E1 ubiquitin-activating like enzyme) and Atg10 (E2 ubiquitin-activating like enzyme). Non-covalent interaction between the Atg12-Atg5 conjugate and Atg16L1, allows for the formation of a large complex which associates with the nascent phagophore. The Atg16L1 complex dissociates from the autophagosome once it is fully formed (1,2).

In the context of its role in autophagy, Atg5 plays diverse physiologically relevant roles. For example, Atg5 together with Atg7 are required for adipogenesis (3). Recently, Atg5 has been implicated in the process of B-cell receptor polarization and antigen presentation (4). In addition to its role in autophagy, Atg5 is implicated in apoptotic cell death. Interaction of Atg5 with FADD (Fas-associated protein with death domain) is involved in cell death induced by IFN-gamma. A truncated form of Atg5, a 24kDa fragment, leads to cell death by interacting with Bcl-xl and inhibiting its anti-apoptotic activity (5). Other Atg5 interacting partners include interleukin-beta (IL-beta) converting enzyme and nucleotide binding oligomerization domain protein 1, which suggest that Atg5 may play other biologically relevant roles (3).

References

1. Yang, Z., & Klionsky, D. J. (2010). Mammalian autophagy: Core molecular machinery and signaling regulation. Current Opinion in Cell Biology. https://doi.org/10.1016/j.ceb.2009.11.014

2. Rubinsztein, D. C., Shpilka, T., & Elazar, Z. (2012). Mechanisms of autophagosome biogenesis. Current Biology. https://doi.org/10.1016/j.cub.2011.11.034

3. Subramani, S., & Malhotra, V. (2013). Non-autophagic roles of autophagy-related proteins. EMBO Reports. https://doi.org/10.1038/embor.2012.220

4. Arbogast, F., Arnold, J., Hammann, P., Kuhn, L., Chicher, J., Murera, D., Gros, F. (2019). ATG5 is required for B cell polarization and presentation of particulate antigens. Autophagy. https://doi.org/10.1080/15548627.2018.1516327

5. Luo, S., & Rubinsztein, D. C. (2007). Atg5 and Bcl-2 provide novel insights into the interplay between apoptosis and autophagy. Cell Death and Differentiation. https://doi.org/10.1038/sj.cdd.4402149

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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Species: Hu
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NB110-53818
Species: Hu, Mu, Rt, Po, Bv, Fi, Gp, Pm, Xp, Ze
Applications: WB, Simple Western, ELISA, EM, Flow, IB, ICC/IF, IHC, IHC-P, IP, PLA, RIA, KD, KO

Publications for ATG5 Antibody (NB110-53818)(212)

We have publications tested in 10 confirmed species: Human, Mouse, Rat, Alligator, Bovine, Drosophila, Guinea Pig, Porcine, Primate, Zebrafish.

We have publications tested in 11 applications: ELISA, EM, Flow, ICC/IF, IHC, IHC-P, IP, KO, PLA, RIA, WB.


Filter By Application
ELISA
(1)
EM
(1)
Flow
(1)
ICC/IF
(13)
IHC
(8)
IHC-P
(9)
IP
(2)
KO
(1)
PLA
(1)
RIA
(1)
WB
(145)
All Applications
Filter By Species
Human
(69)
Mouse
(75)
Rat
(9)
Alligator
(1)
Bovine
(5)
Drosophila
(1)
Guinea Pig
(1)
Porcine
(1)
Primate
(2)
Zebrafish
(9)
All Species
Showing Publications 1 - 10 of 212. Show All 212 Publications.
Publications using NB110-53818 Applications Species
Kern A, Bekbulat F, Schmitt D et al. RAB18 impacts autophagy via lipid droplet-derived lipid transfer and is rescued by ATG9A. bioRxiv Sep 19 2018 (WB, Human) WB Human
Cui L, Song W, Zeng Y et al. Deubiquitinase USP7 regulates Drosophila aging through ubiquitination and autophagy Aging (Albany NY) Nov 20 2020 [PMID: 33221768] (WB, Drosophila) WB Drosophila
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
Her JY, Lee Y, Kim SJ et al. Blockage of protease-activated receptor 2 exacerbates inflammation in high-fat environment partly through autophagy inhibition Aging (Albany NY) Oct 24 2020 [PMID: 33146548] (IP, ICC/IF, Human) IP, ICC/IF Human
Lee DH, Park SH, Ahn J et al. Mir214-3p and Hnf4a/Hnf4 alpha reciprocally regulate Ulk1 expression and autophagy in nonalcoholic hepatic steatosis Autophagy Oct 20 2020 [PMID: 33078654] (WB, Human) WB Human
Park SH, Choi HI, Ahn J et al. Autophagy Functions to Prevent Methylglyoxal-Induced Apoptosis in HK-2 Cells Oxid Med Cell Longev Jun 4 2020 [PMID: 32566104] (WB, Human) WB Human
Mei J Strategy of Hepatic Metabolic Defects Induced by beclin1 Heterozygosity in Adult Zebrafish Int J Mol Sci Feb 24 2020 [PMID: 32102330] (WB, Zebrafish) WB Zebrafish
Marsh T, Kenific CM, Suresh D et al. Autophagic Degradation of NBR1 Restricts Metastatic Outgrowth during Mammary Tumor Progression Dev. Cell Feb 12 2020 [PMID: 32084360] (WB, Human, Mouse) WB Human, Mouse
Hale A, Merchant M, White M Detection and analysis of autophagy in the American alligator (Alligator mississippiensis) J. Exp. Zool. B Mol. Dev. Evol. Feb 15 2020 [PMID: 32061056] (IHC-P, Alligator) IHC-P Alligator
Show All 212 Publications.

Reviews for ATG5 Antibody (NB110-53818) (7) 4.17

Average Rating: 4.1
(Based on 7 reviews)
We have 7 reviews tested in 3 species: Human, Mouse, Rat.

Reviews using NB110-53818:
Filter by Applications
WB
(6)
IHC-P
(1)
All Applications
Filter by Species
Human
(1)
Mouse
(4)
Rat
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot ATG5 NB110-53818
Enlarge
4
reviewed by:
Anonymous
WB Mouse 09/15/2020
View

Summary

ApplicationWestern Blot
Sample TestedLiver tissue
SpeciesMouse
LotI-1
  4
reviewed by:
Anonymous
WB Mouse 03/31/2018
View

Summary

ApplicationWestern Blot
Sample TestedMouse Colon
SpeciesMouse
LotM-1
Western Blot ATG5 NB110-53818
Enlarge
4
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
Enlarge
4
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
Enlarge
4
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
Enlarge
4
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
  5
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 canine, 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. The arrow in the data sheet points to the ATG5/ATG12 band of 56 kDa. There is also a band of 33 kDa which I guess is ATG5? Could you explain the meaning of detecting the ATG5/ATG12 conjugate in preference to ATG5?  Does the antibody 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 bonds ATG12 in a ubiquitous reaction. The lysate that was used was meant to illustrate the ability to detect autophagy by Western blot via ATG5/ATG12. It is possible to detect free 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 does not detect ATG12, which runs at about 23 kDa.
  4. Does this antibody detect free ATG5 monomer in HeLa or HEK 293 cells? 
    • This antibody is able to detect ATG5 (unconjugated form) at ~33 kDa and the ATG12-conjugated form at ~56 kDa. In the Western blot image we have on our website, we show the antibody's ability to detect the autophagy induced conjugate ATG5/ATG12.  A customer review image shows both the conjugated and unconjugated ATG5 in rat cardiomyocytes. Several  publications show this antibody working in Western blot with mouse and human samples. We are not sure if in any of the publications HeLa or HEK 293 cells specifically were used, but if the ATG5 protein is present, this product will 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 tissue sections.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB110-53818AF350
Alexa Fluor 405 NB110-53818AF405
Alexa Fluor 488 NB110-53818AF488
Alexa Fluor 532 NB110-53818AF532
Alexa Fluor 594 NB110-53818AF594
Alexa Fluor 647 NB110-53818AF647
Alexa Fluor 700 NB110-53818AF700
Alexa Fluor 750 NB110-53818AF750
Biotin NB110-53818B
DyLight 350 NB110-53818UV
DyLight 405 NB110-53818V
DyLight 488 NB110-53818G
DyLight 550 NB110-53818R
DyLight 594 NB110-53818DL594
DyLight 650 NB110-53818C
DyLight 755 NB110-53818IR
FITC NB110-53818F
HRP NB110-53818H

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Blogs on ATG5. Showing 1-10 of 32 blog posts - Show all blog posts.

Liver ASK1 activates autophagy to protect against hepatic fat accumulation, non-alcoholic steatohepatitis and fibrosis
By Jamshed Arslan, Pharm. D., PhD. The most common chronic liver disorder worldwide is non-alcoholic fatty liver disease (NAFLD). This obesity-linked disorder can manifest as hepatic fat accumulation (steatosis) wit...  Read full blog post.


  Read full blog post.


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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.

Showing 1-10 of 32 blog posts - Show all blog posts.
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Recent Reviews

4.1
5
1
4
6
3
0
2
0
1
0

Anonymous
09/15/2020
Application: WB
Species: Mouse

 
Anonymous
03/31/2018
Application: WB
Species: Mouse

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

Bioinformatics

Gene Symbol ATG5