Anti-Caspase-9 抗体 [EPR18108] (ab185719)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18108] to Caspase-9
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Caspase-9 antibody [EPR18108]
Caspase-9 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR18108] to Caspase-9 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IPmore details -
種交差性
交差種: Mouse, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: HeLa whole cell lysates untreated and treated with staurosporine 1uM for 4 hours; NIH/3T3 whole cell lysates untreated and treated with staurosporine1uM for 4 hours; Human fetal brain, fetal heart, fetal kidney and fetal liver and pancreas lysates; Mouse heart and kidney lysates. IP: HeLa whole cell lysate treated with staurosporine 1uM for 4 hours.
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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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% 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
EPR18108 -
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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Assay kits
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Isotype control
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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ab185719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 |
1/1000. Detects a band of approximately 46, 37, 35 kDa (predicted molecular weight: 46 kDa).
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IP |
1/40.
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特記事項 |
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WB
1/1000. Detects a band of approximately 46, 37, 35 kDa (predicted molecular weight: 46 kDa). |
IP
1/40. |
ターゲット情報
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機能
Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).
Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9. -
組織特異性
Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes. -
配列類似性
Belongs to the peptidase C14A family.
Contains 1 CARD domain. -
発生段階
Expressed at low levels in fetal heart, at moderate levels in neonate heart, and at high levels in adult heart. -
翻訳後修飾
Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events. - Information by UniProt
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参照データベース
- Entrez Gene: 842 Human
- Entrez Gene: 12371 Mouse
- Omim: 602234 Human
- SwissProt: P55211 Human
- SwissProt: Q8C3Q9 Mouse
- Unigene: 329502 Human
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別名
- APAF-3 antibody
- APAF3 antibody
- Apoptosis related cysteine peptidase antibody
see all
画像
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : CASP9 knockout THP-1 cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Caspase-9 antibody [EPR18108] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab185719 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image, wild-type and CASP9 knockout THP-1 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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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Caspase-9 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab185719 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab185719 was shown to specifically react with Caspase-9 when Caspase-9 knockout samples were used. Wild-type and Caspase-9 knockout samples were subjected to SDS-PAGE. ab185719 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/10000 dilution
Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with staurosporine 1uM for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 35,37,46 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/2500 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate treated with staurosporine 1uM for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 37,39,49 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/5000 dilution
Lane 1 : Human pancreas tissue lysate
Lane 2 : Human fetal liver tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution
Lane 1 : Mouse heart tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 37,39,49 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Caspase-9 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with staurosporine 1uM for 4 hours with ab185719 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab185719 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate treated with staurosporine 10 µg (Input).
Lane 2: ab185719 IP in HeLa whole cell lysate treated with staurosporine.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185719 in HeLa whole cell lysate treated with staurosporine.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (8)
ab185719 は 8 報の論文で使用されています。
- Wu Q et al. Epigallocatechin gallate enhances human lens epithelial cell survival after UVB irradiation via the mitochondrial signaling pathway. Mol Med Rep 25:N/A (2022). PubMed: 35039875
- Ayesha M et al. MiR-4521 plays a tumor repressive role in growth and metastasis of hepatocarcinoma cells by suppressing phosphorylation of FAK/AKT pathway via targeting FAM129A. J Adv Res 36:147-161 (2022). PubMed: 35127170
- Wang JF et al. SP1-induced overexpression of LINC00520 facilitates non-small cell lung cancer progression through miR-577/CCNE2 pathway and predicts poor prognosis. Hum Cell 34:952-964 (2021). PubMed: 33728585
- Huang Y et al. Dehydrocorydaline inhibits the tumorigenesis of breast cancer MDA-MB-231 cells. Mol Med Rep 22:43-50 (2020). PubMed: 32377708
- Yagensky O et al. Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity. Elife 8:N/A (2019). PubMed: 31453805
- Wang X et al. Silence of lncRNA ANRIL represses cell growth and promotes apoptosis in retinoblastoma cells through regulating miR-99a and c-Myc. Artif Cells Nanomed Biotechnol 47:2265-2273 (2019). PubMed: 31184221
- Zhong X et al. Downregulation of microRNA-34a inhibits oxidized low-density lipoprotein-induced apoptosis and oxidative stress in human umbilical vein endothelial cells. Int J Mol Med 42:1134-1144 (2018). PubMed: 29750293
- Luff SA et al. Role of p53 and transcription-independent p53-induced apoptosis in shear-stimulated megakaryocytic maturation, particle generation, and platelet biogenesis. PLoS One 13:e0203991 (2018). PubMed: 30231080