重组Anti-Bid抗体[Y8] (ab32060)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y8] to Bid
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Bid抗体[Y8]
参阅全部 Bid 一抗 -
描述
兔单克隆抗体[Y8] to Bid -
宿主
Rabbit -
特异性
This antibody does not cross-react with other Bcl-2 family members. -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Bid aa 50-150. The exact sequence is proprietary.
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阳性对照
- Jurkat cell lysate and human prostate carcinoma.
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常规说明
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
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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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y8 -
同种型
IgG -
研究领域
相关产品
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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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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32060于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB | (3) |
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
1/50.
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ICC/IF |
1/100.
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说明 |
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Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
1/50. |
ICC/IF
1/100. |
靶标
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功能
The major proteolytic product p15 BID allows the release of cytochrome c (By similarity). Isoform 1, isoform 2 and isoform 4 induce ICE-like proteases and apoptosis. Isoform 3 does not induce apoptosis. Counters the protective effect of Bcl-2. -
组织特异性
Isoform 2 and isoform 3 are expressed in spleen, bone marrow, cerebral and cerebellar cortex. Isoform 2 is expressed in spleen, pancreas and placenta (at protein level). Isoform 3 is expressed in lung, pancreas and spleen (at protein level). Isoform 4 is expressed in lung and pancreas (at protein level). -
结构域
Intact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family. -
翻译后修饰
TNF-alpha induces a caspase-mediated cleavage of p22 BID into a major p15 and minor p13 and p11 products.
Phosphorylated upon DNA damage, probably by ATM or ATR.
p15 BID is ubiquitinated by ITCH; ubiquitination results in proteasome-dependent degradation. -
细胞定位
Cytoplasm; Cytoplasm. Mitochondrion membrane. When uncleaved, it is predominantly cytoplasmic; Mitochondrion membrane. A significant proportion of isoform 2 localizes to mitochondria, it may be cleaved constitutively; Mitochondrion membrane. Associated with the mitochondrial membrane and Mitochondrion membrane. Translocates to mitochondria as an integral membrane protein. - Information by UniProt
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数据库链接
- Entrez Gene: 637 Human
- Omim: 601997 Human
- SwissProt: P55957 Human
- Unigene: 591054 Human
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别名
- Apoptic death agonist antibody
- Apoptotic death agonist BID antibody
- BH3 interacting domain death agonist antibody
see all
图片
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Anti-Bid antibody [Y8] (ab32060) at 1/1000 dilution + Jurkat cell lysate
Predicted band size: 22 kDa
Observed band size: 22 kDa -
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using ab32060 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemistry analysis of MCF7 (human breast adenocarcinoma epithelial cell) labeling Bid with purified ab32060 at 1/500 dilution (10 µg/ml). Cells were fixed with 100% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control: PBS instead of the primary antibody. -
Purified ab32060 at 1/50 dilution (2µg) immunoprecipitating Bid in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab32060 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32060 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 22 kDa -
Overlay histogram showing HeLa cells stained with ab32060 (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 (ab32060, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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All lanes : Anti-Bid antibody [Y8] (ab32060) at 1/500 dilution
Lanes 1-2 : Whole cell lysate prepared from breast cancer clinical extract
Lane 3 : Whole cell lysate prepared from MCF7 cells
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-rabbit IgG-HRP at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 25 minutes
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (25)
ab32060 被引用在 25 文献中.
- Zheng K et al. Effects of targeted Notch1 silencing on the biological processes of the T24 and 5637 cells in vitro. Oncol Lett 21:305 (2021). PubMed: 33732381
- Qu Y et al. The neuroprotection of deproteinized calf blood extractives injection against Alzheimer's disease via regulation of Nrf-2 signaling. Aging (Albany NY) 13:11150-11169 (2021). PubMed: 33819182
- Vernooij L et al. High-Throughput Screening Identifies Idasanutlin as a Resensitizing Drug for Venetoclax-Resistant Neuroblastoma Cells. Mol Cancer Ther 20:1161-1172 (2021). PubMed: 33850004
- Zhang G et al. Identification of a Novel Defined Immune-Autophagy-Related Gene Signature Associated With Clinical and Prognostic Features of Kidney Renal Clear Cell Carcinoma. Front Mol Biosci 8:790804 (2021). PubMed: 34988121
- Gao J et al. Inhibition of esophageal-carcinoma cell proliferation by genistein via suppression of JAK1/2-STAT3 and AKT/MDM2/p53 signaling pathways. Aging (Albany NY) 12:6240-6259 (2020). PubMed: 32276266