重组Anti-Caspase-8抗体[E7] (ab32397)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E7] to Caspase-8
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Caspase-8抗体[E7]
参阅全部 Caspase-8 一抗 -
描述
兔单克隆抗体[E7] to Caspase-8 -
宿主
Rabbit -
特异性
The antibody should recognize both pro-form (55kDa) and p18 cleaved-form of Caspase-8. -
经测试应用
适用于: WBmore details
不适用于: Flow Cyt or ICC/IF -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Caspase-8 aa 200-300 (N terminal). The exact sequence is proprietary.
Database link: Q14790 -
阳性对照
- WB: Wild-type/Wild-type treated with Staurosporin HAP1, Jurkat, SH-SY5Y, IM-9, and HeLa (ab150035) whole cell lysates.
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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: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E7 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32397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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WB | (2) |
1/500 - 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
For unpurified use at 1/500. |
说明 |
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WB
1/500 - 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). For unpurified use at 1/500. |
靶标
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功能
Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-
-AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex. -
组织特异性
Isoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle. -
疾病相关
Defects in CASP8 are the cause of caspase-8 deficiency (CASP8D) [MIM:607271]. CASP8D is a disorder resembling autoimmune lymphoproliferative syndrome (ALPS). It is characterized by lymphadenopathy, splenomegaly, and defective CD95-induced apoptosis of peripheral blood lymphocytes (PBLs). It leads to defects in activation of T-lymphocytes, B-lymphocytes, and natural killer cells leading to immunodeficiency characterized by recurrent sinopulmonary and herpes simplex virus infections and poor responses to immunization. -
序列相似性
Belongs to the peptidase C14A family.
Contains 2 DED (death effector) domains. -
结构域
Isoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex. -
翻译后修饰
Generation of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events.
Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 841 Human
- Omim: 601763 Human
- SwissProt: Q14790 Human
- Unigene: 599762 Human
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别名
- ALPS2B antibody
- Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein antibody
- Apoptosis related cysteine peptidase antibody
see all
图片
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All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : CASP8 knockout HCT 116 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : CASP8 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDaWestern blot: Anti-CASP8 antibody [E7] (ab32397) staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32397 was shown to bind specifically to CASP8. A band was observed at 55 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CASP8 knockout cell line. To generate this image, wild-type and CASP8 knockout HCT 116 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-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CASP8 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDaLanes 1- 4: Merged signal (red and green). Green - ab32397 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32397 was shown to react with Caspase-8 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264958 (knockout cell lysate ab256857) was used. Wild-type HeLa and CASP8 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Wild-type HAP1 whole cell lysate treated with Staurosporin
Lane 3 : Caspase-8 knockout HAP1 whole cell lysate
Lane 4 : Caspase-8 knockout HAP1 whole cell lysate treated with Staurosporin
Lysates/proteins at 20 µg per lane.
Predicted band size: 55 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32397 observed at 55, 43/41 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab32397 was shown to specifically react with HAP1 + Staurosproin when HAP1 + Staurosproin knockout samples were used. Wild-type and HAP1 + Staurosproin knockout samples were subjected to SDS-PAGE. Ab32397 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 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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All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : IM-9 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 55 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution (unpurified) + HeLa cell lysate at 10 µg
Predicted band size: 55 kDa
Observed band size: 55 kDa
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 (32)
ab32397 被引用在 32 文献中.
- Yang J et al. TAK1 Improves Cognitive Function via Suppressing RIPK1-Driven Neuronal Apoptosis and Necroptosis in Rats with Chronic Hypertension. Aging Dis 14:1799-1817 (2023). PubMed: 37196118
- Feng C et al. Excessive activation of IL-33/ST2 in cancer-associated fibroblasts promotes invasion and metastasis in ovarian cancer. Oncol Lett 23:158 (2022). PubMed: 35399326
- Zhao Q et al. The central role of a two-way positive feedback pathway in molecular targeted therapies-mediated pyroptosis in anaplastic thyroid cancer. Clin Transl Med 12:e727 (2022). PubMed: 35184413
- An W et al. Alpinia katsumadai Hayata induces growth inhibition and autophagy‑related apoptosis by regulating the AMPK and Akt/mTOR/p70S6K signaling pathways in cancer cells. Oncol Rep 48:N/A (2022). PubMed: 35730618
- Sun J et al. The lncRNA CASC2 Modulates Hepatocellular Carcinoma Cell Sensitivity and Resistance to TRAIL Through Apoptotic and Non-Apoptotic Signaling. Front Oncol 11:726622 (2021). PubMed: 35145900