Recombinant Anti-SESN2/Sestrin-2 antibody [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, Flow Cyt, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-SESN2/Sestrin-2 antibody [EPR18907]
See all SESN2/Sestrin-2 primary antibodies -
Description
Rabbit monoclonal [EPR18907] to SESN2/Sestrin-2 -
Host species
Rabbit -
Tested Applications & Species
Application Species Flow Cyt MouseHumanICC/IF HumanIP HumanWB MouseRatHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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. ICC/IF: HCT 116 cells. Flow Cyt: NIH/3T3 and HCT 116 cells. IP: HeLa whole cell lysate.
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General notes
This product was previously labelled as SESN2
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18907 -
Isotype
IgG -
Research areas
Associated products
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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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KO cell pellets
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab178518 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Mouse
Human
|
ICC/IF |
Human
|
IP |
Human
|
WB |
Mouse
Rat
Human
|
Application | Abreviews | Notes |
---|---|---|
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. |
|
Flow Cyt |
1/60.
|
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IP |
1/30.
|
Notes |
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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. |
Flow Cyt
1/60. |
IP
1/30. |
Target
- Information by UniProt
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Database links
- 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
see all -
Alternative names
- DKFZp761M0212 antibody
- DKFZp761M02121 antibody
- Hi95 antibody
see all
Images
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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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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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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.
Protocols
Datasheets and documents
Certificate of Compliance
References (6)
ab178518 has been referenced in 6 publications.
- 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
- Wang LX et al. Sestrin2 protects dendritic cells against endoplasmic reticulum stress-related apoptosis induced by high mobility group box-1 protein. Cell Death Dis 11:125 (2020). PubMed: 32071292
- Lees MJ et al. Novel Essential Amino Acid Supplements Following Resistance Exercise Induce Aminoacidemia and Enhance Anabolic Signaling Irrespective of Age: A Proof-of-Concept Trial. Nutrients 12:N/A (2020). PubMed: 32664648
- Carr M et al. Upregulated expression of the antioxidant sestrin 2 identified by transcriptomic analysis of Japanese encephalitis virus-infected SH-SY5Y neuroblastoma cells. Virus Genes 55:630-642 (2019). PubMed: 31292858
- Wang Z et al. Alleviation of sepsis-induced cardiac dysfunction by overexpression of Sestrin2 is associated with inhibition of p-S6K and activation of the p-AMPK pathway. Mol Med Rep 20:2511-2518 (2019). PubMed: 31524263
- Yang K et al. Sestrin2 Suppresses Classically Activated Macrophages-Mediated Inflammatory Response in Myocardial Infarction through Inhibition of mTORC1 Signaling. Front Immunol 8:728 (2017). WB ; Mouse . PubMed: 28713369