Anti-CD63 抗体 [EPR5702] - Late Endosome Marker (ab134045)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5702] to CD63 - Late Endosome Marker
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-CD63 antibody [EPR5702] - Late Endosome Marker
CD63 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR5702] to CD63 - Late Endosome Marker -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-Pmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide. 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: Human platelet, HL-60 and A375 cell lysates and human melanoma tissue lysate. Brefeldin A treated wild-type HAP1 whole cell lysate. IHC-P: Human melanoma and tonsil tissues.
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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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, 59% PBS -
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
EPR5702 -
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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Positive Controls
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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ab134045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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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 30-65 kDa (predicted molecular weight: 26 kDa).
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IHC-P | (1) |
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
特記事項 |
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WB
1/1000 - 1/10000. Detects a band of approximately 30-65 kDa (predicted molecular weight: 26 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
ターゲット情報
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機能
This antigen is associated with early stages of melanoma tumor progression. May play a role in growth regulation. -
組織特異性
Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages. -
配列類似性
Belongs to the tetraspanin (TM4SF) family. -
細胞内局在
Cell membrane. Lysosome membrane. Late endosome membrane. Also found in Weibel-Palade bodies of endothelial cells. Located in platelet dense granules. - Information by UniProt
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参照データベース
- Entrez Gene: 967 Human
- Omim: 155740 Human
- SwissProt: P08962 Human
- Unigene: 445570 Human
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別名
- Lysosomal associated membrane protein 3 antibody
- CD 63 antibody
- CD63 antibody
see all
画像
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All lanes : Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/1000 dilution
Lane 1 : Wild-type HepG2 Vehicle Control Brefeldin A (0 u/mL, 24h) cell lysate
Lane 2 : Wild-type HepG2 Treated Brefeldin A (10 u/mL, 24h) cell lysate
Lane 3 : CD63 knockout HepG2 Vehicle Control Brefeldin A (0 u/mL, 24h) cell lysate
Lane 4 : CD63 knockout HepG2 Treated Brefeldin A (10 u/mL, 24h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?Anti-CD63 antibody [EPR5702] (ab134045) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134045 was shown to bind specifically to CD63. A band was observed at 30 kDa in wild-type HepG2 cell lysates with no signal observed at this size in CD63 knockout cell line ab280796 (knockout cell lysate ab283828). To generate this image, wild-type and CD63 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Brefeldin A treated wild-type HAP1 whole cell lysate
Lane 3 : CD63 knockout HAP1 whole cell lysate
Lane 4 : Brefeldin A treated CD63 knockout HAP1 whole cell lysate
Lane 5 : HL60 whole cell lysate
Lane 6 : Human platelets whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 26 kDaLanes 1 - 6: Merged signal (red and green). Green - ab134045 observed at 26 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab134045 was shown to specifically react with CD63 in wild-type HAP1 Brefeldin A treated cells as signal was lost in HAP1 Brefeldin A treated CD63 knockout cells. Wild-type and CD63 knockout samples were subjected to SDS-PAGE. Ab134045 and ab9484 (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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD63 with purified ab134045 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/5000 dilution (purified)
Lane 1 : A375 whole cell lysate
Lane 2 : HL-60 whole cell lysate
Lane 3 : Human melanoma tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 30-65 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling CD63 with unpurified ab134045 at a dilution of 1/250.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue (negative control) labelling CD63 with purified ab134045 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-CD63 antibody [EPR5702] - Late Endosome Marker (ab134045) at 1/1000 dilution (unpurified)
Lane 1 : Human platelet lysate
Lane 2 : Human melanoma lysate
Lane 3 : HL-60 lysate
Lane 4 : A375 lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 26 kDa
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 (269)
ab134045 は 269 報の論文で使用されています。
- Ye T et al. Large extracellular vesicles secreted by human iPSC-derived MSCs ameliorate tendinopathy via regulating macrophage heterogeneity. Bioact Mater 21:194-208 (2023). PubMed: 36101856
- Wang C et al. A light-activated magnetic bead strategy utilized in spatio-temporal controllable exosomes isolation. Front Bioeng Biotechnol 10:1006374 (2022). PubMed: 36147530
- Shen Z et al. Small extracellular vesicles of hypoxic endothelial cells regulate the therapeutic potential of adipose-derived mesenchymal stem cells via miR-486-5p/PTEN in a limb ischemia model. J Nanobiotechnology 20:422 (2022). PubMed: 36153544
- Hu Q et al. Emodin attenuates severe acute pancreatitis-associated acute lung injury by suppressing pancreatic exosome-mediated alveolar macrophage activation. Acta Pharm Sin B 12:3986-4003 (2022). PubMed: 36213542
- Xu Y et al. Differential expression of plasma exosomal microRNA in severe acute pancreatitis. Front Pharmacol 13:980930 (2022). PubMed: 36249739