Anti-SQSTM1 / p62抗体- Autophagosome Marker (ab56416)
Key features and details
- Mouse monoclonal to SQSTM1 / p62 - Autophagosome Marker
- Suitable for: IHC-P, WB, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
概述
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产品名称
Anti-SQSTM1 / p62抗体- Autophagosome Marker
参阅全部 SQSTM1 / p62 一抗 -
描述
小鼠单克隆抗体to SQSTM1 / p62 - Autophagosome Marker -
宿主
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB Human
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免疫原
Recombinant full length protein corresponding to Human SQSTM1/ p62 aa 1-440.
Database link: Q13501 -
阳性对照
- WB: HeLa, Hap1. Flow Cyt: HeLa. IHC-P: Human lymph node. ICC/IF: HeLa
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常规说明
This product was changed from ascites to tissue culture supernatant on 28th May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.40
Constituent: PBS -
Concentration information loading... -
纯度
Tissue culture supernatant -
克隆
单克隆 -
同种型
IgG2a -
轻链类型
kappa -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab56416 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| 应用 | Species |
|---|---|
| Flow Cyt |
Human
|
| ICC/IF |
Human
|
| IHC-P |
Human
|
| WB |
Human
|
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-P | (3) |
Use at an assay dependent concentration.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
| WB | (22) |
Use at an assay dependent concentration.
|
| ICC/IF | (4) |
Use at an assay dependent concentration.
Fix cells for 15 minutes with 2 mL of 4% paraformaldehyde solution (pH 7.4 with NaOH in PBS). Permeabilize cells by incubating for 15 minutes on ice with 2 mL of 0.1% Triton X-100 in PBS. |
| Flow Cyt |
Use at an assay dependent concentration.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody |
| 说明 |
|---|
|
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
|
WB
Use at an assay dependent concentration. |
|
ICC/IF
Use at an assay dependent concentration. Fix cells for 15 minutes with 2 mL of 4% paraformaldehyde solution (pH 7.4 with NaOH in PBS). Permeabilize cells by incubating for 15 minutes on ice with 2 mL of 0.1% Triton X-100 in PBS. |
|
Flow Cyt
Use at an assay dependent concentration. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody |
靶标
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功能
Adapter protein which binds ubiquitin and may regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin-1. May play a role in titin/TTN downstream signaling in muscle cells. May regulate signaling cascades through ubiquitination. Adapter that mediates the interaction between TRAF6 and CYLD (By similarity). May be involved in cell differentiation, apoptosis, immune response and regulation of K(+) channels. -
组织特异性
Ubiquitously expressed. -
疾病相关
Defects in SQSTM1 are a cause of Paget disease of bone (PDB) [MIM:602080]. PDB is a metabolic bone disease affecting the axial skeleton and characterized by focal areas of increased and disorganized bone turn-over due to activated osteoclasts. Manifestations of the disease include bone pain, deformity, pathological fractures, deafness, neurological complications and increased risk of osteosarcoma. PDB is a chronic disease affecting 2 to 3% of the population above the age of 40 years. -
序列相似性
Contains 1 OPR domain.
Contains 1 UBA domain.
Contains 1 ZZ-type zinc finger. -
结构域
The UBA domain binds specifically 'Lys-63'-linked polyubiquitin chains of polyubiquitinated substrates. Mediates the interaction with TRIM55.
The OPR domain mediates homooligomerization and interactions with PRKCZ, PRKCI, MAP2K5 and NBR1.
The ZZ-type zinc finger mediates the interaction with RIPK1. -
翻译后修饰
Phosphorylated. May be phosphorylated by PRKCZ (By similarity). Phosphorylated in vitro by TTN. -
细胞定位
Cytoplasm. Late endosome. Nucleus. Sarcomere (By similarity). In cardiac muscles localizes to the sarcomeric band (By similarity). Localizes to late endosomes. May also localize to the nucleus. Accumulates in neurofibrillary tangles and in Lewy bodies of neurons from individuals with Alzheimer and Parkinson disease respectively. Enriched in Rosenthal fibers of pilocytic astrocytoma. In liver cells, accumulates in Mallory bodies associated with alcoholic hepatitis, Wilson disease, indian childhood cirrhosis and in hyaline bodies associated with hepatocellular carcinoma. - Information by UniProt
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数据库链接
- Entrez Gene: 8878 Human
- Omim: 601530 Human
- SwissProt: Q13501 Human
- Unigene: 709030 Human
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别名
- A170 antibody
- DMRV antibody
- EBI 3 associated protein of 60 kDa antibody
see all
图片
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Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)All lanes : Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SQSTM1 knockout HEK293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 62 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab56416 observed at 62 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab56416 was shown to react with SQSTM1/p62 in wild-type HEK293T cells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type HEK293T and SQSTM1 knockout HEK293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab56416 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)Monoclonal antibody to SQSTM1 (ab56416) on HeLa cell, antibody concentration 10 ug/ml.
This image was generated using the ascites version of the product.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)ab56416 (1µg/ml) staining SQSTM1 in human lymph node using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.This image was generated using the ascites version of the product.
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Flow Cytometry - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)Overlay histogram showing HeLa cells stained with ab56416 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56416, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This image was generated using the ascites version of the product.
实验方案
文献 (499)
ab56416 被引用在 499 文献中.
- Zhang J et al. Combination therapy with ropivacaine-loaded liposomes and nutrient deprivation for simultaneous cancer therapy and cancer pain relief. Theranostics 10:4885-4899 (2020). PubMed: 32308756
- Zhang L et al. Ethionine Suppresses Mitochondria Autophagy and Induces Apoptosis via Activation of Reactive Oxygen Species in Neural Tube Defects. Front Neurol 11:242 (2020). PubMed: 32318018
- Zhu N et al. Berberine Protects Against Simulated Ischemia/Reperfusion Injury-Induced H9C2 Cardiomyocytes Apoptosis In Vitro and Myocardial Ischemia/Reperfusion-Induced Apoptosis In Vivo by Regulating the Mitophagy-Mediated HIF-1a/BNIP3 Pathway. Front Pharmacol 11:367 (2020). PubMed: 32292345
- Chen X et al. miR-155 facilitates calcium oxalate crystal-induced HK-2 cell injury via targeting PI3K associated autophagy. Exp Mol Pathol 115:104450 (2020). PubMed: 32417393
- Buch BT et al. Colchicine treatment impairs skeletal muscle mitochondrial function and insulin sensitivity in an age-specific manner. FASEB J 34:8653-8670 (2020). PubMed: 32372536
