製品の概要
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製品名Anti-RAB27A antibody
RAB27A 一次抗体 製品一覧 -
製品の詳細Mouse monoclonal to RAB27A
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由来種Mouse
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アプリケーション適用あり: WB, IHC-P, IP, IHC-FoFr, ICC/IFmore details
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種交差性交差種: Mouse, Rat, Human
交差が予測される動物種: Rabbit
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免疫原
Recombinant fragment: YCENPDIVLC GNKSDLEDQR VVKEEEAIAL AEKYGIPYFE TSAANGTNIS QAIEMLLDLI MKRMERCVDK SWIPEGVVRS NGHASTDQLS EEKEKGACGC , corresponding to amino acids 122-222 of Human RAB27A
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特記事項
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab201960 as a replacement.
製品の特性
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製品の状態Liquid
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保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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バッファーPreservative: None
PBS, pH 7.2 -
Concentration information loading... -
精製度Protein G purified
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ポリ/モノモノクローナル
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アイソタイプIgG1
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軽鎖の種類kappa
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研究分野
関連製品
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Compatible Secondaries
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Immunohistochemistry kits
アプリケーション
Our Abpromise guarantee covers the use of ab55667 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 |
|---|---|---|
| WB | Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 25 kDa. | |
| IHC-P | Use a concentration of 3 µg/ml. | |
| IP | Use at an assay dependent concentration. See Abreview. | |
| IHC-FoFr | Use at an assay dependent concentration. PubMed: 21832089 | |
| ICC/IF | 1/200. |
ターゲット情報
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機能Plays a role in cytotoxic granule exocytosis in lymphocytes. Required for both granule maturation and granule docking and priming at the immunologic synapse.
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組織特異性Found in all the examined tissues except in brain. Low expression was found in thymus, kidney, muscle and placenta. Detected in melanocytes, and in most tumor cell lines examined. Expressed in cytotoxic T-lymphocytes (CTL) and mast cells.
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関連疾患Defects in RAB27A are a cause of Griscelli syndrome type 2 (GS2) [MIM:607624]. Griscelli syndrome is a rare autosomal recessive disorder that results in pigmentary dilution of the skin and hair, the presence of large clumps of pigment in hair shafts, and an accumulation of melanosomes in melanocytes. GS2 patients also develop an uncontrolled T-lymphocyte and macrophage activation syndrome, known as hemophagocytic syndrome, leading to death in the absence of bone marrow transplantation. Neurological impairment is present in some patients, likely as a result of hemophagocytic syndrome.
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配列類似性Belongs to the small GTPase superfamily. Rab family.
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細胞内局在Membrane. Melanosome. Late endosome. Lysosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Localizes to endosomal exocytic vesicles.
- Information by UniProt
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参照データベース
- Entrez Gene: 5873 Human
- Entrez Gene: 11891 Mouse
- Entrez Gene: 50645 Rat
- Omim: 603868 Human
- SwissProt: P51159 Human
- SwissProt: Q9ERI2 Mouse
- SwissProt: P23640 Rat
- Unigene: 654978 Human
see all -
別名
- GS2 antibody
- GTP-binding protein Ram antibody
- HsT18676 antibody
see all
画像
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Western blot - Anti-RAB27A antibody (ab55667)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: RAB27A knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jurkat whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab55667 observed at 27 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab55667 was shown to specifically react with RAB27A when RAB27A knockout samples were used. Wild-type and RAB27A knockout samples were subjected to SDS-PAGE. Ab55667 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 2.5 ug/ml and 1/10000 dilution respectively. Blots were developed with IRDye® 800CW Goat anti-Mouse IgG (H + L) and IRDye® 680 Goat anti-Rabbit IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-RAB27A antibody (ab55667)Anti-RAB27A antibody (ab55667) + HL-60 cell lysate
Predicted band size: 25 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-RAB27A antibody (ab55667)This image is courtesy of an anonymous abreview.Immunocytochemistry/ Immunofluorescence analysis of SK-MEL-28 cells labeling RAB27A with ab55667 at 1/200 dilution. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton-X in PBS. A polyclonal goat anti-mouse IgG Alexa Fluor® 488 secondary antibody was used at 1/1000 dilution.
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Western blot - Anti-RAB27A antibody (ab55667)All lanes : Anti-RAB27A antibody (ab55667)
Lane 1 : RAB27A over-expressed 293 cell line, cotransfected with RAB27A Validated Chimera RNAi
Lane 2 : Non-transfected control
Predicted band size: 25 kDaGAPDH (36.1 kDa) used as specificity and loading control.
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Western blot - Anti-RAB27A antibody (ab55667)All lanes : Anti-RAB27A antibody (ab55667)
Lane 1 : RAB27A transfected lysate(24.9 KDa)
Lane 2 : Non-transfected lysate
Predicted band size: 25 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB27A antibody (ab55667)RAB27A antibody (ab55667) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human lymphoma. Heat induced antigen retrieval with Citrate buffer pH 6 has been performed prior to immunostaining. -
Immunocytochemistry/ Immunofluorescence - Anti-RAB27A antibody (ab55667)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovicab55667 staining cow Retinal Pigment Epithelium (RPE) cells by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 10% serum for 20 minutes at 25°C. The primary antibody was diluted 1/200 and incubated with the sample for 12 hours at 4°C. An Alexa Fluor® 546 conjugated goat anti-mouse antibody was used as the secondary. Rab27a, which localizes to the apical site of the RPE cells, is labeled with red fluorescence. Nuclei were counterstained with DAPI (blue).
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Western blot - Anti-RAB27A antibody (ab55667)This image is courtesy of an Abreview submitted by Dr Vladimir MilenkovicAll lanes : Anti-RAB27A antibody (ab55667) at 1/200 dilution
Lane 1 : whole cell lysate of HEK 293 transfected with empty pCDNA3 vector
Lane 2 : whole cell lysate of HEK 293 cells expressing Rab27a-GFP
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-mouse
Developed using the ECL technique.
Predicted band size: 25 kDa
Observed band size: 52 kDa (why is the actual band size different from the predicted?)
Exposure time: 30 minutesRab27a (25kDa)+GFP(27kDa)=52kDa
The blot was blocked with 5% milk for 30 minutes at 25°C and incubated with the primary antibody for 12 hours at 4°C.
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Immunoprecipitation - Anti-RAB27A antibody (ab55667)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovicab55667 (undiluted) was incubated with whole cell lysate of HEK 293 cells transfected with Rab27a-GFP and a Protein G matrix for 12 hours at 4°C to achieve immunoprecipitation. 20µg of lysate were present in the input.
ab55667 (diluted 1/200) was also used in the western blotting step.
Lane 1. lysate of HEK 293 cells expressing Rab27a-GFP
Lane 2. IP with ab55667
Lane 3. Not bound fraction
プロトコール
参考文献
This product has been referenced in:
- Chen L et al. HCC-derived exosomes elicit HCC progression and recurrence by epithelial-mesenchymal transition through MAPK/ERK signalling pathway. Cell Death Dis 9:513 (2018). Read more (PubMed: 29725020) »
- Jiang C et al. GGPP-Mediated Protein Geranylgeranylation in Oocyte Is Essential for the Establishment of Oocyte-Granulosa Cell Communication and Primary-Secondary Follicle Transition in Mouse Ovary. PLoS Genet 13:e1006535 (2017). Read more (PubMed: 28072828) »
レビューと Q&A
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