Anti-RAB10 抗体 [MJF-R23] (ab237703)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MJF-R23] to RAB10
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-RAB10 antibody [MJF-R23]
RAB10 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [MJF-R23] to RAB10 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IP, ICC/IF, WBmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: A549, HeLa, HCT 116, MCF7, NIH/3T3, PC-12 and C6 whole cell lysates. ICC/IF: A549 and MCF7 cells. Flow: A549 cells. IP: A549 whole cell lysate.
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特記事項
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.
This antibody was developed with support from The Michael J. Fox Foundation.
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 long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
MJF-R23 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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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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Recombinant Protein
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Related Products
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ab237703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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Flow Cyt (Intra) |
1/600.
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IP |
1/30.
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ICC/IF |
1/500.
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WB |
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
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特記事項 |
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Flow Cyt (Intra)
1/600. |
IP
1/30. |
ICC/IF
1/500. |
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa). |
ターゲット情報
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機能
The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different set of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion (By similarity). That Rab is mainly involved in the biosynthetic transport of proteins from the Golgi to the plasma membrane. Regulates, for instance, SLC2A4/GLUT4 glucose transporter-enriched vesicles delivery to the plasma membrane. In parallel, it regulates the transport of TLR4, a toll-like receptor to the plasma membrane and therefore may be important for innate immune response. Plays also a specific role in asymmetric protein transport to the plasma membrane within the polarized neuron and epithelial cells. In neurons, it is involved in axonogenesis through regulation of vesicular membrane trafficking toward the axonal plasma membrane while in epithelial cells, it regulates transport from the Golgi to the basolateral membrane. Moreover, may play a role in the basolateral recycling pathway and in phagosome maturation. According to PubMed:23263280, may play a role in endoplasmic reticulum dynamics and morphology controlling tubulation along microtubules and tubules fusion. -
配列類似性
Belongs to the small GTPase superfamily. Rab family. -
細胞内局在
Cytoplasmic vesicle membrane. Golgi apparatus membrane. Golgi apparatus, trans-Golgi network membrane. Endosome membrane. Recycling endosome membrane. Cytoplasmic vesicle, phagosome membrane. Cell projection, cilium. Endoplasmic reticulum membrane. Associates with SLC2A4/GLUT4 storage vesicles (PubMed:22908308). Localizes to the base of the cilium (PubMed:20576682). Transiently associates with phagosomes (By similarity). Localizes to the endoplasmic reticulum at domains of new tubule growth (PubMed:23263280). - Information by UniProt
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参照データベース
- Entrez Gene: 10890 Human
- Entrez Gene: 19325 Mouse
- Entrez Gene: 50993 Rat
- Omim: 612672 Human
- SwissProt: P61026 Human
- SwissProt: P61027 Mouse
- SwissProt: P35281 Rat
- Unigene: 467960 Human
see all -
別名
- GTP binding protein RAB10 antibody
- Rab10 antibody
- RAB10 member RAS oncogene family antibody
see all
画像
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All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : RAB10 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab237703 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab237703 was shown to recognize RAB10 in wild-type A549 cells as signal was lost at the expected MW in RAB10 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RAB10 knockout samples were subjected to SDS-PAGE. Ab237703 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized RAB10 KO A549 (RAB10 knockout human lung carcinoma epithelial cell) (ab261868) labelling RAB10 (red) with ab237703 at 0.04 μg/ml, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/1000 dilution. The nuclear counter stain is DAPI (blue).
Confocal image showing cytoplasmic staining in wild-type A549 cell line, and no staining in RAB10 KO A549 cell line. Images were acquired on the Perkin Elmer® Operetta HCA and a maximum intensity projection of 7 confocal planes is shown for the representative images.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue).
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All lanes : Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/1000 dilution
Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell line) whole cell lysate
Lane 2 : Rab10 knockout A549 whole cell lysate
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell line) whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 7 : C6 (rat glial tumor glial cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 10 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
The WT and Rab10 KO A549 lysates were kindly provided by our collaborator Dr. Dario Alessi, University of Dundee.
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Anti-RAB10 antibody [MJF-R23] (ab237703) at 1/10000 dilution + HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 180 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
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RAB10 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma cell line) whole cell lysate using ab237703 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab237703 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution was used for detection.
Lane 1: A549 whole cell lysate 10μg (input)
Lane 2: ab237703 IP in A549 whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab237703 in A549 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in MCF7 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 (green) with ab237703 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Confocal image showing cytoplasmic staining in A549 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) cells labeling RAB10 with ab237703 at 1/600 dilution (red) compared with the rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% saponin permeabilized A549 wild-type and knock-out cells labeling RAB10 (red) with ab237703 at 0.5 μg/ml, followed by anti-Rabbit secondary at 1/1000 dilution. The nuclear counter stain is DAPI (blue).
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (14)
ab237703 は 14 報の論文で使用されています。
- Liu Z et al. α-Synuclein-containing erythrocytic extracellular vesicles: essential contributors to hyperactivation of monocytes in Parkinson's disease. J Neuroinflammation 19:53 (2022). PubMed: 35193594
- Kumar R et al. A cell-based GEF assay reveals new substrates for DENN domains and a role for DENND2B in primary ciliogenesis. Sci Adv 8:eabk3088 (2022). PubMed: 35196081
- Petropoulou-Vathi L et al. Distinct profiles of LRRK2 activation and Rab GTPase phosphorylation in clinical samples from different PD cohorts. NPJ Parkinsons Dis 8:73 (2022). PubMed: 35676398
- Khan TG et al. The small GTPase RAB10 regulates endosomal recycling of the LDL receptor and transferrin receptor in hepatocytes. J Lipid Res 63:100248 (2022). PubMed: 35753407
- Salvany S et al. Microglial recruitment and mechanisms involved in the disruption of afferent synaptic terminals on spinal cord motor neurons after acute peripheral nerve injury. Glia 69:1216-1240 (2021). PubMed: 33386754