製品の概要
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製品名Anti-Src antibody [Clone 327]
Src 一次抗体 製品一覧 -
製品の詳細Mouse monoclonal [Clone 327] to Src
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由来種Mouse
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特異性Reacts with both v-Src and c-Src proteins.
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アプリケーション適用あり: Flow Cyt, WB, IPmore details
適用なし: ICC/IF,IHC-Fr or IHC-P -
種交差性交差種: Mouse, Rat, Human
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免疫原
Full length native protein (purified) corresponding to Src. Antibody generated by immunizing BALB/c mice with full length native protein (purified) and then fusing with P3X63 Ag8.653 myeloma cells.
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エピトープEpitope is within the SH3 domain of either v-Src or c-Src. We have no evidence for ab16885 cross-reacting with other src family kinases (e.g.fyn).
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ポジティブ･コントロール
- WB: MCF7, HeLa, A431 and NIH/3T3 cell lysates. Flow Cyt: SH-SY5Y cells.
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特記事項
May appear as a doublet due to phosphorylation. May be used to precipitate active Src which can then be used in a kinase reaction.
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 ab222220 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. Store at +4°C. Do Not Freeze.
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バッファーPreservative: 0.1% Sodium azide
Constituents: 0.2% Gelatin, 0.82% Sodium phosphate -
Concentration information loading... -
精製度IgG fraction
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一次抗体 備考May appear as a doublet due to phosphorylation. May be used to precipitate active Src which can then be used in a kinase reaction.
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ポリ/モノモノクローナル
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クローン名Clone 327
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ミエローマP3x63-Ag8.653
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アイソタイプIgG1
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研究分野
関連製品
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Compatible Secondaries
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Conjugation kits
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Isotype control
アプリケーション
Our Abpromise guarantee covers the use of ab16885 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 |
|---|---|---|
| Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| WB | Use a concentration of 2.5 µg/ml. Predicted molecular weight: 59.7 kDa. ab16885 may appear as a doublet on WB due to phosphorylation. This antibody can be used to precipitate active src kinase. | |
| IP | Use at 5 µg/mg of lysate. See Abreview. |
ターゲット情報
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機能Non-receptor protein tyrosine kinase that plays pivotal roles in numerous cellular processes such as proliferation, migration, and transformation. In concert with PTK2B, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. Once it is recruited to the activated integrins, by PTK2B, it phosphorylates CBL which in turn induces the activation and recruitment of phosphatidylinositol 3-kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. Promotes energy production in osteoclasts by activating mitochondrial cytochrome C oxidase. Phosphorylates RUNX3 and COX2 on tyrosine residues, TNK2 on 'Tyr-284' and CBL on 'Tyr-731'. Enhances DDX58/RIG-I-elicited antiviral signaling.
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配列類似性Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain. -
翻訳後修飾Dephosphorylated at Tyr-530 by PTPRJ (By similarity). Phosphorylated on Tyr-530 by c-Src kinase (CSK). The phosphorylated form is termed pp60c-src. Dephosphorylated by PTPRJ at Tyr-419. Normally maintained in an inactive conformation with the SH2 domain engaged with Tyr-530, the SH3 domain engaged with the SH2-kinase linker, and Tyr-419 dephosphorylated. Dephosphorylation of Tyr-530 as a result of protein tyrosine phosphatase (PTP) action disrupts the intramolecular interaction between the SH2 domain and Tyr-530, Tyr-419 can then become autophosphorylated, resulting in SRC activation. Phosphorylation of Tyr-530 by CSK allows this interaction to reform, resulting in SRC inactivation.
S-nitrosylation is important for activation of its kinase activity. -
細胞内局在Cell membrane. Mitochondrion inner membrane.
- Information by UniProt
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参照データベース
- Entrez Gene: 6714 Human
- Entrez Gene: 20779 Mouse
- Entrez Gene: 83805 Rat
- Omim: 190090 Human
- SwissProt: P12931 Human
- SwissProt: P05480 Mouse
- SwissProt: Q9WUD9 Rat
- Unigene: 195659 Human
see all -
製品の状態This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
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別名
- ASV antibody
- Avian sarcoma virus antibody
- c SRC antibody
see all
画像
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![Western blot - Anti-Src antibody [Clone 327] (ab16885)](http://a.abcam.com/ps/products/16/ab16885/Images/ab16885-253297-ab16885KOWB.jpg)
Western blot - Anti-Src antibody [Clone 327] (ab16885)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Src knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab16885 observed at 60 kDa. Red - loading control, ab8227, observed at 42 kDa.ab16885 was shown to specifically react with Src when Src knockout samples were used. Wild-type and Src knockout samples were subjected to SDS-PAGE. ab16885 and ab8227 (loading control to beta Actin) were diluted 2.5µg/ml and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat 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.
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![Flow Cytometry - Anti-Src antibody [Clone 327] (ab16885)](http://a.abcam.com/ps/products/16/ab16885/Images/Src-Primary-antibodies-ab16885-5.jpg)
Flow Cytometry - Anti-Src antibody [Clone 327] (ab16885)Overlay histogram showing SH-SY5Y cells stained with ab16885 (red line). The cells were fixed with 80% methanol (5 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 (ab16885, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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![Western blot - Anti-Src antibody [Clone 327] (ab16885)](http://a.abcam.com/ps/products/16/ab16885/Images/ab16885-277467-Srcab16885web.jpg)
Western blot - Anti-Src antibody [Clone 327] (ab16885)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Src knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab16885 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
This western blot image is a comparison between ab16885 and a competitor's top cited rabbit polyclonal antibody.
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![Western blot - Anti-Src antibody [Clone 327] (ab16885)](http://a.abcam.com/ps/products/16/ab16885/Images/ab16885_review_12897_2.jpg)
Western blot - Anti-Src antibody [Clone 327] (ab16885)This image is courtesy of an anonymous AbreviewAll lanes : Anti-Src antibody [Clone 327] (ab16885) at 2.5 µg/ml
Lane 1 : Whole cell lysate prepared from human MCF-7 cells (Untreated)
Lane 2 : Whole cell lysate prepared from human MCF-7 cells (Treated with 10uM PP2)
Secondary
All lanes : HRP conjugated Donkey polyclonal to mouse Ig at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 59.7 kDa
Exposure time: 3 minutes
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![Western blot - Anti-Src antibody [Clone 327] (ab16885)](http://a.abcam.com/ps/products/16/ab16885/Images/Src-Primary-antibodies-ab16885-4.jpg)
Western blot - Anti-Src antibody [Clone 327] (ab16885)This image is a courtesy of Anonymous AbreviewAnti-Src antibody [Clone 327] (ab16885) at 1/100 dilution + Lysate prepared from mouse NIH 3T3 cell line at 10 µg
Secondary
HRP-conjugated sheep monoclonal to mouse IgG at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 59.7 kDa
Observed band size: 60 kDa (why is the actual band size different from the predicted?)
Exposure time: 12 minutes
参考文献
This product has been referenced in:
- Zhou J et al. Black rice-derived anthocyanins inhibit HER-2-positive breast cancer epithelial-mesenchymal transition-mediated metastasis in vitro by suppressing FAK signaling. Int J Mol Med 40:1649-1656 (2017). Read more (PubMed: 29039492) »
- Albanito L et al. Effects of atrazine on estrogen receptor a- and G protein-coupled receptor 30-mediated signaling and proliferation in cancer cells and cancer-associated fibroblasts. Environ Health Perspect 123:493-9 (2015). Read more (PubMed: 25616260) »
レビューと Q&A
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"