Anti-Rab9 抗体 [Mab9] (ab2810)
Key features and details
- Mouse monoclonal [Mab9] to Rab9
- Suitable for: WB, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Hamster, Cow, Dog, Human
- Isotype: IgG1
製品の概要
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製品名
Anti-Rab9 antibody [Mab9]
Rab9 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [Mab9] to Rab9 -
由来種
Mouse -
アプリケーション
適用あり: WB, Flow Cyt, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Hamster, Cow, Dog, Human
交差が予測される動物種: Non human primates -
免疫原
Recombinant full length protein corresponding to Dog Rab9.
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ポジティブ・コントロール
- WB: MDA-MB-231, MCF7, HeLa, HEK293, Jurkat, HepG2 and K562 cell lysates. Flow Cyt: THP1 cells. IHC-P: Human tonsil and spleen tissues.
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特記事項
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as 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. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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精製度
IgG fraction -
一次抗体 備考
Rab proteins are a family of Ras-like GTPases involved in intracellular compartment protein transport. Different members of the 40+ member rab family are responsible for docking and fusion of transport vesicles between different compartments within the cell. Rab 9 is required for trafficking mannose 6-phosphate receptor between the late endosome to trans-Golgi network (TGN). By facilitating receptor transport, rab 9 enables cells to efficiently recycle important cellular trafficking components. It is functionally necessary for rab 9, like other rab family members, to be prenylated by two 20-carbon geranylgeranyl groups at the C-terminus. Most prenylated rab 9 is membrane bound, however, 10-20% of the cellular pool of rab 9 is bound to GDP dissociation inhibitor-alpha (GDI-alpha) in the cytosol. GDI recycles prenylated, GDP bound rab 9 from their fusion targets back to their membranes of origin. -
ポリ/モノ
モノクローナル -
クローン名
Mab9 -
アイソタイプ
IgG1 -
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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Positive Controls
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Recombinant Protein
アプリケーション
Our Abpromise guarantee covers the use of ab2810 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 特記事項 |
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WB | Use a concentration of 10 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa). | |
Flow Cyt | 1/100. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ターゲット情報
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機能
Involved in the transport of proteins between the endosomes and the trans Golgi network. -
配列類似性
Belongs to the small GTPase superfamily. Rab family. -
細胞内局在
Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 511776 Cow
- Entrez Gene: 403947 Dog
- Entrez Gene: 9367 Human
- Entrez Gene: 56382 Mouse
- Entrez Gene: 84589 Rat
- Omim: 300284 Human
- SwissProt: P24408 Dog
- SwissProt: P51151 Human
see all -
別名
- 2410064E05Rik antibody
- AI195561 antibody
- DmRab9 antibody
see all
画像
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All lanes : Anti-Rab9 antibody [Mab9] (ab2810) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RAB9A knockout HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : MDA-MB-231 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
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 - ab2810 observed at 25 kDa. Red - loading control ab181602 observed at 36 kDa.
ab2810 Anti-Rab9 antibody [Mab9] was shown to specifically react with Rab9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265693 (knockout cell lysate ab257625) was used. Wild-type and Rab9 knockout samples were subjected to SDS-PAGE. ab2810 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 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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All lanes : Anti-Rab9 antibody [Mab9] (ab2810) at 10 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 5 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 22 kDa
Additional bands at: 48 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab9 antibody [Mab9] (ab2810)This image is courtesy of an Abreview submitted by Qin Wen
ab2810 staining Rab9 in Mouse adult testes tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 4% paraformaldehyde and blocked with 5% BSA for 60 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in 1% BSA) for 15 hours at 4°C. A TRITC-conjugated Donkey anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.
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Overlay histogram showing THP1 cells stained with ab2810 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab2810, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab9 antibody [Mab9] (ab2810)ab2810 (2µg/ml) staining Rab9 in human spleen, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic and cell membrane staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab9 antibody [Mab9] (ab2810)IHC image of Rab9 staining in human normal tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2810, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
参考文献 (21)
ab2810 は 21 報の論文で使用されています。
- Kim WY et al. Atg7-dependent canonical autophagy regulates the degradation of aquaporin 2 in prolonged hypokalemia. Sci Rep 9:3021 (2019). PubMed: 30816234
- Saito T et al. An alternative mitophagy pathway mediated by Rab9 protects the heart against ischemia. J Clin Invest 129:802-819 (2019). PubMed: 30511961
- Kucera A et al. Spatiotemporal Resolution of Rab9 and CI-MPR Dynamics in the Endocytic Pathway. Traffic 17:211-29 (2016). PubMed: 26663757
- Tsai SY et al. Regulation of TLR3 Activation by S100A9. J Immunol 195:4426-37 (2015). PubMed: 26385519
- Zhou GL et al. Deficits in receptor-mediated endocytosis and recycling in cells from mice with Gpr107 locus disruption. J Cell Sci 127:3916-27 (2014). PubMed: 24849652