Anti-APG5L/ATG5 抗体 [EPR1755(2)] - BSA and Azide free (ab221604)
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 - BSA and Azide free
- Suitable for: ICC/IF, IP, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-APG5L/ATG5 antibody [EPR1755(2)] - BSA and Azide free
APG5L/ATG5 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5 - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, IP, WB, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
ポジティブ・コントロール
- Raji, HeLa, HT-1080, Human fetal kidney, C6, Raw264.7, PC-12 and NIH3T3 cell lysates; Human hepatocellular carcinoma and Human ovarian adenocarcinoma tissue
-
特記事項
ab221604 is the carrier-free version of ab108327.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR1755(2) -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
KO cell lines
-
Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab221604の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
ICC/IF |
Use at an assay dependent concentration.
|
|
IP |
Use at an assay dependent concentration.
|
|
WB |
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.
|
|
IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
特記事項 |
---|
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
ターゲット情報
-
機能
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. -
組織特異性
Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells. -
配列類似性
Belongs to the ATG5 family. -
翻訳後修飾
Conjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane.
Acetylated by EP300. -
細胞内局在
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. - Information by UniProt
-
参照データベース
- 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
see all -
別名
- APG 5 antibody
- APG 5L antibody
- APG5 antibody
see all
画像
-
All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : ATG5 knockout THP-1 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-APG5L/ATG5 antibody [EPR1755(2)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108327 was shown to bind specifically to APG5L/ATG5. A band was observed at 50 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG5 knockout cell line ab277835 (knockout cell lysate ab290722). To generate this image, wild-type and ATG5 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
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 (HRP) (ab131366) was used for detection at 1/1000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108327).
プロトコール
データシートおよび資料
-
Datasheet download
Certificate of Compliance
参考文献 (14)
ab221604 は 14 報の論文で使用されています。
- Li M et al. Erastin triggers autophagic death of breast cancer cells by increasing intracellular iron levels. Oncol Lett 20:57 (2020). PubMed: 32793311
- Wang ZC et al. MicroRNA-137 inhibits autophagy and chemosensitizes pancreatic cancer cells by targeting ATG5. Int J Biochem Cell Biol N/A:N/A (2019). PubMed: 30710750
- Gómez-Puerto MC et al. Autophagy proteins ATG5 and ATG7 are essential for the maintenance of human CD34(+) hematopoietic stem-progenitor cells. Stem Cells N/A:N/A (2016). PubMed: 26930546
- Chen Y et al. FGFR antagonist induces protective autophagy in FGFR1-amplified breast cancer cell. Biochem Biophys Res Commun N/A:N/A (2016). WB ; Human . PubMed: 26993162
- Wu Y et al. Targeting the MIR34C-5p-ATG4B-autophagy axis enhances the sensitivity of cervical cancer cells to pirarubicin. Autophagy N/A:0 (2016). PubMed: 27097054