重组Anti-SESN2/Sestrin-2抗体[EPR18907] (ab178518)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18907] to SESN2/Sestrin-2
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-SESN2/Sestrin-2抗体[EPR18907]
参阅全部 SESN2/Sestrin-2 一抗 -
描述
兔单克隆抗体[EPR18907] to SESN2/Sestrin-2 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IP, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa whole cell lysate treated with 10 mM H2O2 for 1 hour; HeLa, LoVo, 293, NIH/3T3, HCT 116, Rat1, RAW 264.7, C6 and PC-12 whole cell lysates; Human colon, fetal liver, testis and fetal kidney lysates; Mouse spleen lysate. HEK-293 cell lysate. ICC/IF: HCT 116 cells. Flow Cyt (intra): NIH/3T3 and HCT 116 cells. IP: HeLa whole cell lysate.
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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.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
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
EPR18907 -
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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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab178518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 |
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
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ICC/IF |
1/100.
ICC/IF is recommended for human and rat only. |
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IP |
1/30.
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Flow Cyt (Intra) |
1/60.
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说明 |
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WB
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). |
ICC/IF
1/100. ICC/IF is recommended for human and rat only. |
IP
1/30. |
Flow Cyt (Intra)
1/60. |
靶标
- Information by UniProt
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数据库链接
- Entrez Gene: 83667 Human
- Entrez Gene: 230784 Mouse
- Entrez Gene: 502988 Rat
- Omim: 607767 Human
- SwissProt: P58004 Human
- SwissProt: P58043 Mouse
- Unigene: 469543 Human
- Unigene: 23608 Mouse
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别名
- DKFZp761M0212 antibody
- DKFZp761M02121 antibody
- Hi95 antibody
see all
图片
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All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 cell lysate
Lane 2 : SESN2 knockout HEK-293 cell lysate
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 54 kDaFalse colour image of Western blot: Anti-SESN2/Sestrin-2 antibody [EPR18907] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab178518 was shown to bind specifically to SESN2/Sestrin-2. A band was observed at 54 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in SESN2 knockout cell line ab269486 (knockout cell lysate ab269650). To generate this image, wild-type and SESN2 knockout HEK-293 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-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SESN2 knockout HeLa cell lysate
Lane 3 : LoVo cell lysate
Lane 4 : HEK-293T 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
Predicted band size: 54 kDa
Observed band size: 54 kDaLanes 1-4: Merged signal (red and green). Green - ab178518 observed at 54 kDa. Red - loading control ab8245 observed at 36 kDa.
ab178518 Anti-SESN2/Sestrin-2 antibody [EPR18907] was shown to specifically react with SESN2/Sestrin-2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265669 (knockout lysate ab257665) was used. Wild-type and SESN2/Sestrin-2 knockout samples were subjected to SDS-PAGE. ab178518 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling SESN2/Sestrin-2 with ab178518 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HCT116 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/250 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SESN2 knockout HeLa cell lysate
Lane 3 : HeLa 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
Predicted band size: 54 kDa
Observed band size: 54 kDaLanes 1-3: Merged signal (red and green). Green - ab178518 observed at 54 kDa. Red - loading control ab8245 observed at 36 kDa.
ab178518 Anti-SESN2/Sestrin-2 antibody [EPR18907] was shown to specifically react with SESN2/Sestrin-2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265669 (knockout cell lysate ab257665) was used. Wild-type and SESN2/Sestrin-2 knockout samples were subjected to SDS-PAGE. ab178518 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 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 10 mM H2O2 for 1 hour
Lane 3 : LoVo (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 4 : 293T (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Sestrin expression is induced by H2O2 treatment, which is consistent with what has been described in the literature (PMID: 25337554).
Exposure time:3 minutes
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All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution
Lane 1 : Human colon lysate
Lane 2 : Human fetal liver lysate
Lane 3 : Human testis lysate
Lane 4 : Human fetal kidney lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time:3 minutes
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All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution
Lane 1 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
Lane 2 : Rat1 (Rat fibroblast cell line) whole cell lysate
Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : Mouse spleen lysate
Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1, 2, 3, 4 and 5: 3 minutes; Lane 6: 30 seconds.
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SESN2/Sestrin-2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab178518 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab178518 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate, 10µg (Input).
Lane 2: ab178518 IP in HeLa whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab178518 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
SESN2/Sestrin-2 expression is low in HeLa cells and can be enriched through immunoprecipitation.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling SESN2/Sestrin-2 with ab178518 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal[EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HCT 116 (Human colorectal carcinoma cell line) cells labeling SESN2/Sestrin-2 with ab178518 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (14)
ab178518 被引用在 14 文献中.
- Zhang S & Jin Z Bone Mesenchymal Stem Cell-Derived Extracellular Vesicles Containing Long Noncoding RNA NEAT1 Relieve Osteoarthritis. Oxid Med Cell Longev 2022:5517648 (2022). PubMed: 35480871
- Li JY et al. Sestrin2 protects dendrite cells against ferroptosis induced by sepsis. Cell Death Dis 12:834 (2021). PubMed: 34482365
- Zhao Y et al. Exercise May Promote Skeletal Muscle Hypertrophy via Enhancing Leucine-Sensing: Preliminary Evidence. Front Physiol 12:741038 (2021). PubMed: 34630161
- Wang T et al. Exercise improves lipid metabolism disorders induced by high-fat diet in a SESN2/JNK- independent manner. Appl Physiol Nutr Metab N/A:N/A (2021). PubMed: 34038646
- Lin Q et al. Sestrin-2 regulates podocyte mitochondrial dysfunction and apoptosis under high-glucose conditions via AMPK. Int J Mol Med 45:1361-1372 (2020). PubMed: 32323727