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

产品名称

Anti-APG5L/ATG5抗体[EPR1755(2)]
参阅全部 APG5L/ATG5 一抗
描述

兔单克隆抗体[EPR1755(2)] to APG5L/ATG5
宿主

Rabbit
经测试应用

适用于: ICC/IF, WB, IP, IHC-Pmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- Raji, HeLa, HT-1080, Human fetal kidney, C6, Raw264.7, PC-12 and NIH3T3 cell lysates; Human hepatocellular carcinoma and Human ovarian adenocarcinoma tissue
常规说明
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.

形式

Liquid
存放说明

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.
存储溶液

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

Protein A purified
克隆

单克隆
克隆编号

EPR1755(2)
同种型

IgG
研究领域
- Neuroscience
- Processes

Alternative Versions
- HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
- Anti-APG5L/ATG5 antibody [EPR1755(2)] - BSA and Azide free (ab221604)
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

Abpromise™承诺保证使用ab108327于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
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.

说明
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.

功能

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.
组织特异性

Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells.
序列相似性

Belongs to the ATG5 family.
翻译后修饰

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

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
数据库链接
- 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
别名
- 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

Country/Region

Language
Datasheet download

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发表研究结果有使用 ab108327？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab108327 被引用在 131 文献中.

Liang L et al. miR-30d-5p suppresses proliferation and autophagy by targeting ATG5 in renal cell carcinoma. FEBS Open Bio 11:529-540 (2021). PubMed: 33145996
Hoffmann HH et al. TMEM41B Is a Pan-flavivirus Host Factor. Cell 184:133-148.e20 (2021). PubMed: 33338421
Kato M et al. miR-379 deletion ameliorates features of diabetic kidney disease by enhancing adaptive mitophagy via FIS1. Commun Biol 4:30 (2021). PubMed: 33398021
Guo S et al. TRIB2 modulates proteasome function to reduce ubiquitin stability and protect liver cancer cells against oxidative stress. Cell Death Dis 12:42 (2021). PubMed: 33414446
Liu Z et al. Targeting autophagy enhances atezolizumab-induced mitochondria-related apoptosis in osteosarcoma. Cell Death Dis 12:164 (2021). PubMed: 33558476

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1-7 of 7 Abreviews or Q&A

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

提交于 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

提交于 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

提交于 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.

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Verified customer

提交于 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.

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Verified customer

提交于 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

提交于 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

提交于 Mar 24 2014

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (131)

客户评价及客户问答

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)]