AB309155

Anti-Ubiquitin (phospho S65) 抗体 [30H3/30K1]

Anti-Ubiquitin (phospho S65) antibody [30H3/30K1]

RabMAb
Recombinant
詳細を見る

Be the first to review this product! Submit a review

(4 Publications)

Rabbit Recombinant Monoclonal UBC phospho S65 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), I-ELISA and reacts with Human, Mouse, Synthetic peptide - Human samples. Cited in 4 publications.

別名を表示する

Polyubiquitin-C, UBC

6 Images

Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) cells labelling Ubiquitin (phospho S65) with ab309155 at 1/250 dilution (3.652 ug/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic and nuclear staining in PC-3 cells treated with CCCP (30μM) (ab141229) for 6 h. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

Flow Cytometry (Intracellular) - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-3 cells (human pancreas adenocarcinoma epithelial cells) treated with 30μM CCCP (ab141229) for 6 hours (Red) / untreated PC-3 cells (Green) cells labelling Ubiquitin (phospho S65) with ab309155 at 1/1000 dilution (0.1 ug) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Indirect ELISA - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

I-ELISA

Supplier Data

Indirect ELISA - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Indirect ELISA analysis of ab309155 at 0-1000 ng/ml. The secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution. Antigen : Human polyubiquitin-C (phospho S65) peptide1, Human polyubiquitin-C (phospho S65) peptide 2,Human polyubiquitin-C non-phospho peptide. Antigen concentration : 1000 ng/ml

Supplier Data

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, Anti-Ubiquitin antibody (ab134953) staining at 1/1000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 97 seconds.

All lanes:

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (ab309155) at 1/1000 dilution

Lane 1:

Untreated Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEK-293T treated with 20 uM CCCP (<a href='/products/biochemicals/cccp-mitochondrial-oxidative-phosphorylation-uncoupler-ab141229'>ab141229</a>) for 8 hours, whole cell lysate at 20 µg

Lane 3:

Untreated PINK1 knockout HEK-293T whole cell lysate at 20 µg

Lane 4:

PINK1 knockout HEK-293T treated with 20 uM CCCP (<a href='/products/biochemicals/cccp-mitochondrial-oxidative-phosphorylation-uncoupler-ab141229'>ab141229</a>) for 8 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 8 kDa,200 kDa

false

Exposure time: 97s

Supplier Data

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. In Western blot, Anti-Ubiquitin antibody (ab134953) staining at 1/1000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (ab309155) at 1/1000 dilution

Lane 1:

Untreated PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Untreated PC-3 whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 3:

PC-3 treated with 30 uM CCCP (<a href='/products/biochemicals/cccp-mitochondrial-oxidative-phosphorylation-uncoupler-ab141229'>ab141229</a>) for 2 hours, whole cell lysate (untreated membrane) at 20 µg

Lane 4:

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 8 kDa,200 kDa

false

Exposure time: 180s

Lab

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

ab181602 was used as GAPDH loading control.

All lanes:

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (ab309155) at 1/1000 dilution

Lane 1:

Untreated MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 2:

MEF treated with 5 μM Valinomycin for 24 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 26 kDa

Observed band size: 8-200 kDa,27 kDa

false

Exposure time: 26s

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

30H3/30K1

アイソタイプ

IgG

キャリアフリー

交差種

Human, Mouse

アプリケーション

WB, ICC/IF, Flow Cyt (Intra), I-ELISA

applications

免疫原

The exact immunogen used to generate this antibody is proprietary information.

style

left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IELISA" : {"fullname" : "Indirect ELISA", "shortname":"I-ELISA"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/250", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/1000", "FlowCytIntra-species-notes": "", "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "" }, "Synthetic peptide": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IELISA-species-checked": "testedAndGuaranteed", "IELISA-species-dilution-info": "1/1000", "IELISA-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "" } } }

style

left-column

製品の詳細

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Collaborations
This antibody was developed with support from The Michael J. Fox Foundation.

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

style

left-column

出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Protein A

バッファー組成

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

出荷温度

Blue Ice

短期保存期間

1-2 weeks

短期保存温度

+4°C

長期保存温度

-20°C

分注に関する情報

Upon delivery aliquot

保管に関する情報

Avoid freeze / thaw cycle

style

left-column

補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Ubiquitin is a small regulatory protein found in almost all tissues of eukaryotic organisms. It has a molecular weight of approximately 8.6 kilodaltons. It functions mechanically by attaching to proteins through a process called ubiquitination which involves the formation of an isopeptide bond. Ubiquitin molecules can form polyubiquitin chains through different lysine residues such as K48 and K63 that determine their function. These chains label substrate proteins for various fates including degradation. Ubiquitin is expressed ubiquitously in cells reflecting its essential role in maintaining protein homeostasis.

Biological function summary

The ubiquitin system plays a critical role in regulating protein turnover and quality control within cells. It is part of a larger complex known as the ubiquitin-proteasome system (UPS) which is responsible for degrading proteins that need to be turned over. This process is essential for cell cycle control response to oxidative stress and DNA repair. Ubiquitin's role in tagging proteins for degradation or signaling allows cells to respond quickly to changes in their environment and maintain balance.

Pathways

Ubiquitin functions in several important biological pathways including the Wnt and NF-kB pathways. In the Wnt signaling pathway ubiquitination modulates the stability of key components thereby affecting the pathway's overall activity. In the NF-kB signaling pathway ubiquitin labels inhibitor proteins for degradation which releases and activates NF-kB. These pathways highlight ubiquitin's interaction with proteins such as beta-catenin in Wnt and IkB in NF-kB illustrating how it regulates diverse cellular processes.

The dysfunction of the ubiquitin system is linked to neurodegenerative diseases and cancers. Ubiquitin-related defects in protein degradation can lead to the buildup of unwanted proteins contributing to conditions like Parkinson's disease. Connections with cancer are evident as ubiquitin controls cell cycle proteins and aberrant ubiquitination may drive tumor growth and progression. The protein p53 known to be controlled by ubiquitination plays a significant role in cancer related mechanisms when dysregulated. Understanding and targeting ubiquitin-related pathways may provide new therapeutic opportunities for treating these conditions.

style

left-column

style

left-column

製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

style

left-column

ターゲットの情報

Ubiquitin. Exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked : Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in proteotoxic stress response and cell cycle; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. During ubiquitination, the acceptor ubiquitin is positioned in the active site via direct interaction with the E2 ubiquitin-conjugating enzymes such as UBE2R2 (PubMed : 38326650). As a monoubiquitin, its C-terminal glycine is recognized as a C-degron by Cul2-RING (CRL2) E3 ubiquitin-protein ligase complexes (PubMed : 39548056).

See full target information Polyubiquitin-C phospho S65

style

left-column

文献 (4)

Recent publications for all applications. Explore the full list and refine your search

iScience 28:111544 PubMed39811672

2025

ZFAND6 promotes TRAF2-dependent mitophagy to restrain cGAS-STING signaling.

Applications

Unspecified application

Species

Unspecified reactive species

Kashif Shaikh,Melissa Bowman,Sarah M McCormick,Linlin Gao,Jiawen Zhang,Jesse White,John Tawil,Arun Kapoor,Ravit Arav-Boger,Christopher C Norbury,Edward W Harhaj

Autophagy 20:2076-2091 PubMed38802071

2024

Development and characterization of phospho-ubiquitin antibodies to monitor PINK1-PRKN signaling in cells and tissue.

Applications

Unspecified application

Species

Unspecified reactive species

Jens O Watzlawik,Xu Hou,Tyrique Richardson,Szymon L Lewicki,Joanna Siuda,Zbigniew K Wszolek,Casey N Cook,Leonard Petrucelli,Michael DeTure,Dennis W Dickson,Odetta Antico,Miratul M K Muqit,Jordan B Fishman,Karima Pirani,Ravindran Kumaran,Nicole K Polinski,Fabienne C Fiesel,Wolfdieter Springer

Biochimica et biophysica acta. Molecular cell research 1871:119712 PubMed38521466

2024

DDIT3/CHOP promotes LPS/ATP-induced pyroptosis in osteoblasts via mitophagy inhibition.

Applications

Species

Mouse

Zhipeng Dong,Beining Yang,Meie Jia,Chang Yang,Shuo Wang,Hailin Mu,Jiawei Wang

bioRxiv : the preprint server for biology : PubMed38293125

2024

Development and characterization of phospho-ubiquitin antibodies to monitor PINK1-PRKN signaling in cells and tissue.

Applications

WB, ELISA, ICC, WB, ELISA, ICC

Species

Human, Human, Human, Mouse, Mouse, Mouse

View all publications

style

left-column

style

left-column

style

right-column

Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Ubiquitin (phospho S65) 抗体 [30H3/30K1]

Anti-Ubiquitin (phospho S65) antibody [30H3/30K1]

Immunocytochemistry/ Immunofluorescence - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Flow Cytometry (Intracellular) - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Indirect ELISA - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

Western blot - Anti-Ubiquitin (phospho S65) antibody [30H3/30K1] (AB309155)

関連する標識済み抗体及び組成の異なる製品 (1)

Key facts

宿主種

クローン性

クローン番号

アイソタイプ

キャリアフリー

交差種

アプリケーション

免疫原

Reactivity data

製品の詳細

出荷温度及び保存条件

製品の状態

精製方法

バッファー組成

出荷温度

短期保存期間

短期保存温度

長期保存温度

分注に関する情報

保管に関する情報

補足情報

Biological function summary

Pathways

製品プロトコール

ターゲットの情報

文献 (4)

組合せ製品

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Abcam product promise