Name: Anti-APG5L/ATG5 antibody [EPR1755(2)]
Brand: RabMAb
SKU: ab108327
Rating: 4 (7 reviews)

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Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5
Suitable for: ICC/IF, WB, IP, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free HRP

Product name

Anti-APG5L/ATG5 antibody [EPR1755(2)]
See all APG5L/ATG5 primary antibodies
Description

Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, WB, IP, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Raji, HeLa, HT-1080, Human fetal kidney, C6, Raw264.7, PC-12 and NIH3T3 cell lysates; Human hepatocellular carcinoma and Human ovarian adenocarcinoma tissue
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR1755(2)
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human APG5L/ATG5 protein (ab103787)
Related Products
- Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108327 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ICC/IF	(1)	1/100 - 1/250.
WB	(4)	1/1000 - 1/10000. Predicted molecular weight: 32 kDa.
IP		1/10 - 1/100.
IHC-P	(1)	1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended.

Notes
ICC/IF 1/100 - 1/250.
WB 1/1000 - 1/10000. Predicted molecular weight: 32 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended.

Function

Involved in autophagic vesicle formation. Conjugation with ATG12, through a ubiquitin-like conjugating system involving ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3-like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. Involved in mitochondrial quality control after oxidative damage, and in subsequent cellular longevity. The ATG12-ATG5 conjugate also negatively regulates the innate antiviral immune response by blocking the type I IFN production pathway through direct association with RARRES3 and MAVS. Also plays a role in translation or delivery of incoming viral RNA to the translation apparatus. Plays a critical role in multiple aspects of lymphocyte development and is essential for both B and T lymphocyte survival and proliferation. Required for optimal processing and presentation of antigens for MHC II. Involved in the maintenance of axon morphology and membrane structures, as well as in normal adipocyte differentiation. Promotes primary ciliogenesis through removal of OFD1 from centriolar satellites and degradation of IFT20 via the autophagic pathway.
May play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity. Plays a crucial role in IFN-gamma-induced autophagic cell death by interacting with FADD.
Tissue specificity

Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells.
Sequence similarities

Belongs to the ATG5 family.
Post-translational
modifications

Conjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane.
Acetylated by EP300.
Cellular localization

Cytoplasm. Preautophagosomal structure membrane. Colocalizes with nonmuscle actin. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed (By similarity). Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
Target information above from: UniProt accession Q9H1Y0 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 9474 Human
- Entrez Gene: 11793 Mouse
- Entrez Gene: 365601 Rat
- Omim: 604261 Human
- SwissProt: Q9H1Y0 Human
- SwissProt: Q99J83 Mouse
- SwissProt: Q3MQ06 Rat
- Unigene: 486063 Human
- Unigene: 22264 Mouse
- Unigene: 98385 Rat
see all
Alternative names
- APG 5 antibody
- APG 5L antibody
- APG5 antibody
- APG5 autophagy 5 like antibody
- APG5 like antibody
- APG5-like antibody
- APG5L antibody
- Apoptosis specific protein antibody
- Apoptosis-specific protein antibody
- ASP antibody
- ATG 5 antibody
- Atg5 antibody
- ATG5 autophagy related 5 homolog antibody
- ATG5_HUMAN antibody
- Autophagy protein 5 antibody
- Autophagy related 5 antibody
- hAPG5 antibody
- Homolog of S Cerevisiae autophagy 5 antibody
- OTTHUMP00000040507 antibody
see all

Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: APG5L/ATG5 knockout HAP1 cell lysate (20 µg)
Lane 3: Raji cell lysate (20 µg)
Lane 4: Jeg-3 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab108327 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab108327 was shown to specifically react with APG5L/ATG5 when APG5L/ATG5 knockout samples were used. Wild-type and APG5L/ATG5 knockout samples were subjected to SDS-PAGE. ab108327 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human ovarian adenocarcinoma tissue by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Immunofluorescence staining of MCF7 cells with purified ab108327 at a working dilution of 1/150, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108327 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution (purified)

Lane 1 : C6 cell lysate
Lane 2 : PC-12 cell lysate
Lane 3 : NIH/3T3 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution (purified) + Raji cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution (purified) + HT-1080 cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution (purified) + human fetal kidney at 10 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 32 kDa
Observed band size: 32,55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab180327 at a working dilution of 1/150. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunoprecipitation - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

ab180327 (purified) at 1/20 immunoprecipitating CRSP8 in 10 μg PC-12 whole cell lysate (Lanes 1 and 2, observed at 55 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution (unpurified)

Lane 1 : Raji cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HT-1080 cell lysate
Lane 4 : Human fetal kidney cell lysate
Lane 5 : C6 cell lysate
Lane 6 : Raw264.7 cell lysate
Lane 7 : PC-12 cell lysate
Lane 8 : NIH3T3 cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 32 kDa

Secondary antibody - anti-rabbit HRP (ab6721)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human hepatocellular carcinoma tissue by Immunohistochemistry.
Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab108327? Please let us know so that we can cite the reference in this datasheet.

ab108327 has been referenced in 162 publications.

Ji P et al. Genetically engineered probiotics as catalytic glucose depriver for tumor starvation therapy. Mater Today Bio 18:100515 (2023). PubMed: 36582449
Deng Q et al. A detrimental role of NLRP6 in host iron metabolism during Salmonella infection. Redox Biol 49:102217 (2022). PubMed: 34942528
Zhuang A et al. Metformin promotes histone deacetylation of optineurin and suppresses tumour growth through autophagy inhibition in ocular melanoma. Clin Transl Med 12:e660 (2022). PubMed: 35075807
Wu MD et al. Acetylshikonin induces autophagy-dependent apoptosis through the key LKB1-AMPK and PI3K/Akt-regulated mTOR signalling pathways in HL-60 cells. J Cell Mol Med 26:1606-1620 (2022). PubMed: 35106915
Zhang W et al. Autophagic Schwann cells promote perineural invasion mediated by the NGF/ATG7 paracrine pathway in pancreatic cancer. J Exp Clin Cancer Res 41:48 (2022). PubMed: 35109895

View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Baboon Tissue sections (adrenal)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 0.01M citrate buffer (pH 6.0)

Permeabilization

Yes - 0.4% Triton X-100 in KPBS

Specification

adrenal

Blocking step

Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 22°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 29 2022

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (testis cell)

Permeabilization

Specification

testis cell

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 15 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Leukemic Cancer Cell lines)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

25 µg

Specification

Leukemic Cancer Cell lines

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 31 2018

Application

Flow Cytometry

Sample

Human Cell (Peripheral Blood Mononuclear Cells)

Permeabilization

Yes - 90% Methanol on ice, 20min.

Gating Strategy

FSC v SSC to gate out debris and dying cells followed by CD8+ve selection.

Specification

Peripheral Blood Mononuclear Cells

Preparation

Cell harvesting/tissue preparation method: Cells isolated from whole blood using Dextran/Percoll method.
Sample buffer: 2% FCS

Fixation

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 06 2017

Application

Western blot

Sample

Human Cell lysate - whole cell (Hepato carcinoma HepG2 cells)

Gel Running Conditions

Reduced Denaturing (4 - 15%)

Loading amount

20 µg

Specification

Hepato carcinoma HepG2 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 17 2016

Application

Western blot

Loading amount

30 µg

Gel Running Conditions

Reduced Denaturing

Sample

Human Cell lysate - whole cell (macrophage)

Specification

macrophage

Treatment

0.1 ug/ml LPS 24 h

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Dec 12 2014

Application

Western blot

Loading amount

40 µg

Gel Running Conditions

Non-reduced Denaturing (12)

Sample

Xenopus tropicalis Tissue lysate - whole (Brain)

Specification

Brain

Treatment

protease inhibitor cocktail treatment in RIPA lysis buffer

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 24 2014

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (162)

Customer reviews and Q&As

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Western blot abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Flow Cytometry abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Western blot abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Western blot abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Western blot abreview for Anti-APG5L/ATG5 antibody [EPR1755(2)]

Post-translational
modifications