Anti-ZAP70 抗体 [E267] (ab32410)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E267] to ZAP70
- Suitable for: WB
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-ZAP70 antibody [E267]
ZAP70 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [E267] to ZAP70 -
由来種
Rabbit -
特異性
ab32410 recognises Zap-70, a Syk-family protein tyrosine kinase. -
アプリケーション
適用あり: WBmore details
適用なし: ICC/IF or IHC -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Rat -
免疫原
Synthetic peptide within Human ZAP70 aa 300-400 (N terminal). The exact sequence is proprietary.
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ポジティブ・コントロール
- Jurkat whole cell lysate (ab7899)
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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.
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. Avoid freeze / thaw cycles. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 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
E267 -
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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Isotype control
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Positive Controls
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Recombinant Protein
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ab32410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 |
1/500. Predicted molecular weight: 70 kDa.
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特記事項 |
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WB
1/500. Predicted molecular weight: 70 kDa. |
ターゲット情報
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機能
Plays a role in T-cell development and lymphocyte activation. Essential for TCR-mediated IL-2 production. Isoform 1 induces TCR-mediated signal transduction, isoform 2 does not. -
組織特異性
Expressed in T- and natural killer cells. -
関連疾患
Defects in ZAP70 are the cause of selective T-cell defect (STD) [MIM:176947]. STD is an autosomal recessive form of severe combined immunodeficiency characterized by a selective absence of CD8-type T-cells. -
配列類似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily.
Contains 1 protein kinase domain.
Contains 2 SH2 domains. -
ドメイン
The SH2 domains bind to the phosphorylated tyrosine-based activation motif (TAM) of CD3Z and the non-canonical phosphorylated tyrosine-based activation motif (TAM) of RHOH. -
翻訳後修飾
Phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Tyr-319 phosphorylation is essential for full activity. -
細胞内局在
Cytoplasm. Cell membrane. After antigen stimulation, isoform 1 concentrates at the immunological synapse and isoform 2 remains cytoplasmic. Co-localizes together with RHOH in the immunological synapse. RHOH is required for its proper localization to the cell membrane and cytoskeleton fractions in the thymocytes. - Information by UniProt
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参照データベース
- Entrez Gene: 7535 Human
- Entrez Gene: 22637 Mouse
- Entrez Gene: 301348 Rat
- Omim: 176947 Human
- SwissProt: P43403 Human
- SwissProt: P43404 Mouse
- Unigene: 234569 Human
- Unigene: 8038 Mouse
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別名
- 70 kDa zeta associated protein antibody
- 70 kDa zeta-associated protein antibody
- EC 2.7.10.2 antibody
see all
画像
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All lanes : Anti-ZAP70 antibody [E267] (ab32410) at 1/500 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : ZAP70 CRISPR-Cas9 edited Jurkat cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : Raji cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 70 kDaFalse colour image of Western blot: Anti-ZAP70 antibody [E267] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32410 was shown to bind specifically to ZAP70. A band was observed at 70 kDa in wild-type Jurkat cell lysates with no signal observed at this size in ZAP70 CRISPR-Cas9 edited cell line ab273841 (CRISPR-Cas9 edited cell lysate ab273795). The band observed in the CRISPR-Cas9 edited lysate lane below 70 kDa is likely to represent a truncated form of ZAP70. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and ZAP70 CRISPR-Cas9 edited Jurkat 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-ZAP70 antibody [E267] (ab32410) at 1/1000 dilution
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed)
Predicted band size: 70 kDaAnti-GAPDH antibody, ab8245 (1/20000) was used as a primary antibody for the loading control and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed, ab216776 (1/10000) was used as a loading control secondary antibody.
Lanes 1-2: Merged signal (red and green). Green – ab32410 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.
ab32410 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 10000 dilution for 1 hour at room temperature before imaging.The expression profile observed in Raji is consistent with the literature (PMID: 25275600).
Negative control: Raji (PMID: 25275600)
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Anti-ZAP70 antibody [E267] (ab32410) at 1/500 dilution + Jurkat cell lysates
Predicted band size: 70 kDa
Observed band size: 70 kDa
Western blot analysis on Jurkat Cell lysate.
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
参考文献 (3)
ab32410 は 3 報の論文で使用されています。
- Han L et al. IgD-Fc-Ig fusion protein, a new biological agent, inhibits T cell function in CIA rats by inhibiting IgD-IgDR-Lck-NF-?B signaling pathways. Acta Pharmacol Sin 41:800-812 (2020). PubMed: 31937932
- Schleiss C et al. BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo. Sci Rep 9:701 (2019). PubMed: 30679590
- Liang W et al. Core Fucosylation of the T Cell Receptor Is Required for T Cell Activation. Front Immunol 9:78 (2018). PubMed: 29434598