2. 抗体
  3. 一抗
  4. 单克隆抗体 重组抗体
  5. ITCH 抗体 (YA2623)

ITCH Antibody (YA2623) 是一个兔来源、无偶联标记、抗 ITCH 的 IgG 单克隆抗体。

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Top Publications Citing Use of Products
  • WB: 蛋白质免疫印迹;
  • IHC-P: 石蜡切片样本的免疫组织化学;
  • IHC-F: 冰冻切片样本的免疫组织化学;
  • ICC/IF: 细胞免疫荧光;
  • IF-Tissue: 组织免疫荧光;
  • mIHC: 多重荧光免疫组化;
  • IP: 免疫沉淀;
  • ChIP: 染色质免疫沉淀;
  • FC: 流式细胞术;
  • ELISA: 酶联免疫吸附试验

  • 产品详情

  • 验证图片

  • 背景信息

  • 产品资料

描述

ITCH Antibody (YA2623) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ITCH.
宿主

Rabbit
克隆性

Recombinant,Monoclonal
分子量
Predicted band size: 103 kDa;
Observed band size: 103 kDa
请注意：因蛋白存在修饰或聚体等情况，以实测为准，预测仅为参考。
反应种属
Human, Mouse, Rat
蛋白数据库
基因 ID
免疫原

A synthesized peptide derived from human ITCH aa800-850.
应用 & 推荐
稀释比例
应用 稀释比
WB
WB: 蛋白质免疫印迹
1:500-1:1000
IHC-P
IHC-P: 石蜡切片样本的免疫组织化学
1:50-1:100
敏感性 Endogenous 纯度 Affinity Purified
偶联 Non-conjugated 修饰 Unmodified
同型 IgG  
性状

液体
组分

Supplied in rabbit IgG in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40%Glycerol, 0.01% sodium azide and 0.05% BSA.
保存条件 & 期限

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
运输条件

Shipping with blue ice.
验证图片
ALL WB IHC mIHC

  • Western blot analysis was performed on extracts from K562 (lane 1, 15 μg), HeLa (lane 2, 15 μg), 293T (lane 3, 15 μg), Ramos (lane 4, 15 μg), Mouse brain (lane 5, 15 μg), and Rat brain (lane 6, 15 μg) using ITCH/AIP4 Rabbit mAb.Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin, HY-P80438, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

  • Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using ITCH antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated overnight at 4°C with the primary antibody (HY-P82878,1:50 dilution). Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using ITCH antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated overnight at 4°C with the primary antibody (HY-P82878,1:50 dilution). Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using ITCH antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated overnight at 4°C with the primary antibody (HY-P82878,1:50 dilution). Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using ITCH antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated overnight at 4°C with the primary antibody (HY-P82878,1:50 dilution). Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Liver Cancer tissue using ITCH antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated overnight at 4°C with the primary antibody (HY-P82878,1:50 dilution). Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Immunohistochemical analysis of paraffin-embedded human Colon cancer‌ tissue using ITCH antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated overnight at 4°C with the primary antibody (HY-P82878,1:50 dilution). Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Tonsil tissue using ITCH antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82878, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Tonsil tissue using ITCH antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82878, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Tonsil tissue using ITCH antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82878, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Tonsil tissue using ITCH antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82878, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Tonsil tissue using ITCH antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82878, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

  • Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Tonsil tissue using ITCH antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P82878, 1:50 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

背景
功能：作为一种 E3 泛素蛋白连接酶，它从 E2 泛素结合酶接受以硫酯形式存在的泛素，然后直接将泛素转移到靶底物上 (PubMed:11046148, PubMed:14602072, PubMed:15051726, PubMed:16387660, PubMed:17028573, PubMed:18718448, PubMed:18718449, PubMed:19116316, PubMed:19592251, PubMed:19881509, PubMed:20068034, PubMed:20392206)。 PubMed:20491914、PubMed:23146885、PubMed:24790097、PubMed:25631046)。催化 Lys-29、Lys-48 和 Lys-63 连接的泛素化 (PubMed:17028573、PubMed:18718448、PubMed:19131965、PubMed:19881509)。参与炎症信号通路调控 (PubMed:19131965)。 RIPK1 是泛素编辑蛋白复合物的重要组成部分，该复合物还包括 TNFAIP3、TAX1BP1 和 RNF11，确保炎症信号通路的瞬时性 (PubMed:19131965)。TNFAIP3 促进 TNF 刺激后该复合物的形成 (PubMed:19131965)。复合物形成后，TNFAIP3 去除 RIPK1 上的 Lys-63 多聚泛素链，并催化 Lys-48 多聚泛素链的形成 (PubMed:19131965)。这导致 RIPK1 被蛋白酶体降解，从而终止 TNF 或 LPS 介导的 NFKB1 激活 (PubMed:19131965)。通过赖氨酸-63 连接的泛素化修饰 RIPK2，并影响 NOD2 依赖的信号转导通路 (PubMed:19592251)。调节多种转录因子的转录活性，可能在免疫反应的调控中发挥重要作用 (PubMed:18718448, PubMed:20491914)。通过赖氨酸-63 连接的泛素化修饰 NFE2，并参与造血谱系发育的调控 (PubMed:18718448)。介导 JUN 的泛素化和降解 (基于相似性)。介导 JUNB 的泛素化和降解 (PubMed:16387660)。通过诱导 JUNB 泛素化和降解，该蛋白是 2 型辅助性 T 细胞 (Th2) 细胞因子产生的关键调节因子 (基于相似性)。参与 MAVS 依赖性细胞抗病毒反应的负调控 (PubMed:19881509)。通过 Lys-48 连接的泛素化修饰 MAVS，导致 MAVS 被蛋白酶体降解 (PubMed:19881509)。配体刺激后，该蛋白可能通过调节 PI42KA 活性来调控 Wnt 受体 FZD4 向降解性内吞途径的转运 (PubMed:23146885)。该蛋白泛素化 PI4K2A 并负调控其催化活性 (PubMed:23146885)。泛素化趋化因子受体 CXCR4，并通过泛素化转运所需的内体分选复合物 ESCRT-0 组分 HGS 和 STAM，调节 CXCR4 在配体刺激后向降解性内吞途径的转运 (PubMed:14602072, PubMed:23146885, PubMed:34927784)。靶向 DTX1 进行溶酶体降解，并在无配体的情况下，通过 Lys-29 连接的多聚泛素化控制 NOTCH1 的降解 (PubMed:17028573, PubMed:18628966, PubMed:23886940)。泛素化 SNX9 (PubMed:20491914)。 ITCH 通过 Lys-48 连接的泛素化修饰 MAP3K7 (基于相似性)。它与 UBR5 共同参与 TXNIP 的泛素化和蛋白酶体降解，从而调控细胞凋亡和活性氧水平：催化 TXNIP 的 Lys-48/Lys-63 分支泛素化 (PubMed:20068034, PubMed:29378950)。ITCH 合成 Lys-63 连接的泛素链，而 UBR5 则在最初修饰的底物上形成多个 Lys-48 连接的泛素链 (PubMed:29378950)。通过泛素化和蛋白酶体降解 p15 BID 介导表皮生长因子的抗凋亡活性 (PubMed:20392206)。泛素化 BRAT1，且在 NDFIP1 存在下该泛素化作用增强 (PubMed:25631046)。通过 Lys-48 连接的泛素化修饰甲型流感病毒 (IAV) 基质蛋白 1 (M1)，抑制 IAV 的复制，导致 M1 蛋白酶体降解 (PubMed:30328013)。泛素化 NEDD9/HEF1，导致 NEDD9/HEF1 蛋白酶体降解 (PubMed:15051726)。
亚细胞定位：细胞膜；外周膜蛋白；胞质侧；细胞质；细胞核；早期内体膜；外周膜蛋白；胞质侧；内体膜；外周膜蛋白；胞质侧
表达水平：
组织特异性：广泛表达
异构体 & 翻译后修饰：Q96J02 有 3 种异构体：Q96J02-1：102803 Da (预测值)；Q96J02-2：98676 Da (预测值)；Q96J02-3：86498 Da (预测值)。
T 细胞活化后，JNK 级联对 PRR 结构域周围丝氨酸和苏氨酸残基的磷酸化会加速 JUN 和 JUNB 的泛素化和降解。JNK1 磷酸化导致 ITCH 催化活性增强，这可能是由于构象变化破坏了 PRR/WW 基序结构域和 HECT 结构域之间的相互作用，从而使 HECT 结构域暴露出来 (基于相似性推测)。FYN 磷酸化会降低与 JUNB 的相互作用，并负调控 JUN 的泛素化和降解；单泛素化 (PubMed：19116316)。具有“Lys-63”连接的自聚泛素化不会导致蛋白质降解 (PubMed:18718449, PubMed:23146885, PubMed:24790097)
亚基：单体。由 SMAD3、ITCH/AIP4 和 NEDD9/HEF1 组成的三元复合物的一部分；在该复合物中，NEDD9/HEF1 通过 N 端与 ITCH/AIP4 (通过 WW 结构域) 相互作用；该复合物介导 NEDD9/HEF1 的泛素化和蛋白酶体降解 (PubMed:15051726)。通过 WW 结构域与 OCNL 相互作用 (基于相似性)。通过 WW 结构域与 NOTCH1 相互作用 (基于相似性)。通过 WW 结构域与 JUN 相互作用 (基于相似性)。与 JUNB 相互作用；该相互作用促进 ITCH 介导的 JUNB 泛素化和降解 (PubMed:16387660)。与 FYN 相互作用；该相互作用使 ITCH 的 Tyr-420 位点磷酸化，从而降低 JUNB 的结合 (PubMed:16387660)。通过 WW 结构域 2 与 N4BP1 相互作用；该相互作用抑制 E3 泛素连接酶活性 (基于相似性)。与 NDFIP1 和 NDFIP2 相互作用；该相互作用激活 E3 泛素连接酶，并可能诱导其募集至外泌体 (基于相似性)。与 ARHGEF7 相互作用 (PubMed:17652093)。与 RNF11 相互作用 (PubMed:14559117, PubMed:19131965)。通过 WW 结构域 1 与 NFE2 相互作用 (通过 PXY 基序 1)。该相互作用促进 NFE2 的 Lys-63 连接的泛素化，使其滞留在细胞质中并抑制其转录激活活性 (PubMed:11318614, PubMed:18718448)。通过 WW 结构域与 CXCR4 (通过 C 端) 相互作用；该相互作用依赖于 CXCR4 的磷酸化 (PubMed:19116316)。与 E3 连接酶 DTX3L 以及 ESCRT-0 组分 HGS 和 STAM 形成复合物 (PubMed:24790097)。通过 C 端与 DTX3L 相互作用；CXCL12 刺激后该相互作用增强，并抑制 ITCH 的催化活性 (该相互作用是直接的)(PubMed:24790097)。与 HGS 相互作用 (PubMed:14602072)。通过 WW 结构域与 PCBP2 相互作用，形成包含 ITCH、MAVS 和 PCBP2 的复合物 (PubMed:19881509)。通过 WW 结构域 (经由 C 端) 与 TXNIP 相互作用 (PubMed:20068034)。与 p15 BID 相互作用 (PubMed:20392206)。与 ERBB4 相互作用 (PubMed:20858735)。与 DTX1 相互作用 (PubMed:17028573)。与 SPART 相互作用 (PubMed:19580544)。与 SNX9 和 SNX18 相互作用 (PubMed:20491914)。通过其 WW 结构域与 ATN1 相互作用 (PubMed:9647693)。通过 WW 结构域与 SGK3 相互作用 (PubMed:16888620)。与 CBLC 相互作用 (PubMed:12226085)。与 OTUD7B 相互作用 (PubMed:22179831)。通过 WW 结构域 1、2 和 3 与 PI4K2A 相互作用；该相互作用抑制 PI4K2A 的催化活性并促进 ITCH 的催化活性 (PubMed:23146885)。与 ARRDC4 相互作用 (PubMed:23236378)。是包含 ITCH、NDFIP1 和 MAP3K7 的复合物的组成部分 (基于相似性)。与 UBE2L3 相互作用；该相互作用由 NDFIP1 介导 (PubMed:25632008)。与 MAPK8/JNK1 相互作用 (基于相似性)。通过 WW 结构域与 ARRDC1 (通过 PPxY 基序) 相互作用；这种相互作用是直接的，并参与泛素蛋白连接酶 ITCH 向 NOTCH1 受体的募集 (PubMed:21191027, PubMed:23886940)。通过 WW 结构域与 ARRDC2 相互作用 (PubMed:21191027)。通过 WW 结构域与 ARRDC3 相互作用 (PubMed:21191027, PubMed:23886940)。直接与 LDLRAD3 相互作用；这种相互作用促进 ITCH 的自身泛素化，导致其降解 (PubMed:26854353)。与 ENTREP1 相互作用；它能增强 ITCH 对 CXCR4 的泛素化及其后续的内吞作用 (PubMed:34927784)。它与 USP12 和 WDR48/UAF1 相互作用；当 USP12 和 WDR48/UAF1 同时参与时，这种相互作用更有效，并且可能促进 USP12 去泛素化酶复合物募集到 Notch 上 (PubMed:22778262)。
反应种属数据库

Entrez Gene: 83737 Human ; 16396 Mouse ; 311567 Rat

SwissProt: Q96J02 Human ; Q8C863 Mouse ;

OMIM: 613385 Human

研究领域

Cell Biology
中文名
ITCH 抗体 (YA2623)
同用名
ITCH; E3 ubiquitin-protein ligase Itchy homolog; Itch; Atrophin-1-interacting protein 4; AIP4; NFE2-associated polypeptide 1; NAPP1
文件资料

COA (194 KB)

抗体使用指南 (1083 KB)

ITCH Antibody (YA2623) 相关分类

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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