Anti-USP9x 抗体 (ab19879)
Key features and details
- Rabbit polyclonal to USP9x
- Suitable for: ICC/IF, IP, ICC, IHC-Fr, WB, IHC (PFA fixed)
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-USP9x antibody
USP9x 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to USP9x -
由来種
Rabbit -
特異性
ab19879 detects 289kDa full length USP9X Human protein (Q93008) in WB on Caco2 Lysate. All detected bands are quenched by the immunizing peptide ab20617 -
アプリケーション
適用あり: ICC/IF, IP, ICC, IHC-Fr, WB, IHC (PFA fixed)more details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human USP9x.
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ポジティブ・コントロール
- ICC: Hek293 cells
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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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
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ab19879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 |
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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ICC |
Use a concentration of 5 µg/ml.
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IHC-Fr | (1) |
Use at an assay dependent concentration.
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WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 100,105, 290 kDa (predicted molecular weight: 54, 100,105 , 290 kDa).
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IHC (PFA fixed) |
Use a concentration of 0.1 µg/ml.
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特記事項 |
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ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
ICC
Use a concentration of 5 µg/ml. |
IHC-Fr
Use at an assay dependent concentration. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 100,105, 290 kDa (predicted molecular weight: 54, 100,105 , 290 kDa). |
IHC (PFA fixed)
Use a concentration of 0.1 µg/ml. |
ターゲット情報
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機能
Deubiquitinase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. May therefore play an important role regulatory role at the level of protein turnover by preventing degradation of proteins through the removal of conjugated ubiquitin. Essential component of TGF-beta/BMP signaling cascade. Regulates chromosome alignment and segregation in mitosis by regulating the localization of BIRC5/survivin to mitotic centromeres. Specifically hydrolyzes both 'Lys-29'- and 'Lys-33'-linked polyubiquitins chains. Specifically deubiquitinates monoubiquitinated SMAD4, opposing the activity of E3 ubiquitin-protein ligase TRIM33. -
組織特異性
Widely expressed in embryonic and adult tissues. -
配列類似性
Belongs to the peptidase C19 family. -
細胞内局在
Cytoplasm. - Information by UniProt
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参照データベース
- Entrez Gene: 8239 Human
- Entrez Gene: 22284 Mouse
- Entrez Gene: 363445 Rat
- Omim: 300072 Human
- SwissProt: Q93008 Human
- SwissProt: P70398 Mouse
- Unigene: 77578 Human
- Unigene: 242646 Mouse
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別名
- Deubiquitinating enzyme FAF X antibody
- Deubiquitinating enzyme FAF-X antibody
- DFFRX antibody
see all
画像
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ab19879 staining USP9x in Hek293 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab19879 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-USP9x antibody (ab19879) at 1 µg/ml
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : USP9X knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54, 100,105 , 290 kDa
Observed band size: 290 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab19879 observed at 290 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab19879 was shown to react with USP9x in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265665 (knockout cell lysate ab257790) was used. Wild-type HeLa and USP9X knockout HeLa 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. ab19879 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 µg/ml and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Immuofluorescent staining for USP9X in the rat hippocampus (dentate gyrus) using ab19879 (1/300 = 0.07µg/ml). Image is taken with X10 objective. ab19879 was incubated overnight at RT. Secondary antibody used was anti-rabbit Alexa fluor 488 (1/1000 for 2h at RT). Rats were intracardially perfused with paraformaldehyde 4%, brain tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut on a cryostat for free floating IHC.
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USP9x was immunoprecipitated using 0.5mg Caco2 whole cell extract, 5µg of Rabbit polyclonal to USP9x and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Caco2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19879.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 290kDa: USP9x. -
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: USP9x knockout HAP1 cell lysate (20 µg)
Lane 3: T84 cell lysate (20 µg)
Lane 4: NIH3T3 cell lysate (20 µg)
Lanes 1 to 4: Merged signal (red and green). Green - ab19879 observed at 290 kDa. Red - loading control, ab181602 observed at 124 kDa.
ab19879 was shown to specifically react with USP9x when USP9x knockout samples were used. Wild-type and USP9x knockout samples were subjected to SDS-PAGE. ab9879 and ab181602 (loading control to GAPDH) were both diluted at 1 µg/ml and 1/10000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-USP9x antibody (ab19879) at 1 µg/ml
Lane 1 : Caco-2 whole cell lysate (ab3950)
Lane 2 : Caco-2 whole cell lysate (ab3950) with Human USP9x peptide (ab20617) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 54, 100,105 , 290 kDa
Observed band size: 100,105,290 kDa why is the actual band size different from the predicted?
Additional bands at: ~50-65 kDa. We are unsure as to the identity of these extra bands.ab19879 detects full length USP9x protein as well as a number of USP9x fragments in WB on Caco2 Lysate:
289kDa Human protein: Q93008 USP9X (Full length protein)
105kDa Human protein: Q6P468 - USP9X protein (Fragment) Human protein
99.7kDa Human protein: Q59EZ5 - USP9X protein variant (Fragment).
53.9kDa Q86X58 - USP9X protein (Fragment).
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (17)
ab19879 は 17 報の論文で使用されています。
- Deng Y et al. ARV-771 Acts as an Inducer of Cell Cycle Arrest and Apoptosis to Suppress Hepatocellular Carcinoma Progression. Front Pharmacol 13:858901 (2022). PubMed: 35600879
- Xu X et al. Hsa_circ_0008434 regulates USP9X expression by sponging miR-6838-5p to promote gastric cancer growth, migration and invasion. BMC Cancer 21:1289 (2021). PubMed: 34856948
- Wrobel L et al. Transient siRNA-mediated protein knockdown in mouse followed by feeding/starving cycle and liver tissue analysis. STAR Protoc 2:100500 (2021). PubMed: 33997814
- Wrobel L et al. mTORC2 Assembly Is Regulated by USP9X-Mediated Deubiquitination of RICTOR. Cell Rep 33:108564 (2020). PubMed: 33378666
- Yoon S et al. Usp9X Controls Ankyrin-Repeat Domain Protein Homeostasis during Dendritic Spine Development. Neuron 105:506-521.e7 (2020). PubMed: 31813652