AB257665

Human SESN2 (Sestrin-2) knockout HeLa cell lysate

Name: Human SESN2 (Sestrin-2) knockout HeLa cell lysate (ab257665) | Abcam
Brand: Abcam Limited
SKU: AB257665

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SESN2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon4.

View Alternative Names

Hi95, Hypoxia-induced gene, SEST2, SESN2, Sestrin-2

3 Images

Lab

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : SESN2 knockout HeLa cell lysate (20 ug)
Lane 3 : LoVo cell lysate (20 ug)
Lane 4 : HEK-293T cell lysate (20 ug)

ab178518 was shown to specifically react with SESN2/Sestrin-2 in wild-type HeLa cells. Loss of signal was observed when knockout sample 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.

All lanes:

Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] (<a href='/en-us/products/primary-antibodies/sesn2-sestrin-2-antibody-epr18907-ab178518'>ab178518</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SESN2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-sesn2-sestrin-2-knockout-hela-cell-line-ab265669'>ab265669</a>)

Lane 3:

LoVo cell lysate at 20 µg

Lane 4:

HEK-293T cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 38 kDa,54 kDa

Observed band size: 48 kDa,54 kDa

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Lab

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : SESN2 knockout HeLa cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)

ab178518 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^oC 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.

All lanes:

Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] (<a href='/en-us/products/primary-antibodies/sesn2-sestrin-2-antibody-epr18907-ab178518'>ab178518</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SESN2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-sesn2-sestrin-2-knockout-hela-cell-line-ab265669'>ab265669</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

Sanger Sequencing - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Sanger seq

Unknown

Sanger Sequencing - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Homozygous : 1 bp deletion in exon4

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon4.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name

SESN2

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

SESN2 also known as Sestrin-2 or Hi95 is a stress-inducible protein that plays a role in cellular responses to oxidative stress. It has a molecular mass of approximately 60 kDa and is expressed in various tissues including the liver heart and muscle. Sestrin-2 acts as an antioxidant protein that helps reduce reactive oxygen species (ROS) and maintains cellular redox balance. It contains domains that directly interact with target proteins to regulate cellular activity enabling cells to respond efficiently to stress signals.

Biological function summary

Sestrin-2 regulates cell survival and homeostasis during oxidative stress conditions. It modulates the activity of key signaling pathways that control autophagy and cell growth. Sestrin-2 does not function alone but interacts with other proteins to form functional complexes. This allows it to fine-tune cellular processes and contributes to its role in promoting cell repair mechanisms. Its induction during stress conditions demonstrates its engagement in promoting cellular adaptation and resistance to damage.

Pathways

Sestrin-2 engages in signaling networks such as the AMPK and mTOR pathways. Its interaction with AMP-activated protein kinase (AMPK) triggers energy homeostasis and metabolic stress responses. It also negatively regulates the mammalian target of rapamycin (mTOR) to support autophagy and inhibit cell growth when necessary. Sestrin-2 therefore plays an important role in coordinating responses between these pathways ensuring optimal cellular function during periods of environmental stress or nutrient deprivation.

SESN2 is implicated in conditions related to oxidative stress and metabolic dysregulation such as cancer and type 2 diabetes. Its involvement in cancer is connected to its regulatory role on the mTOR pathway often overactive in tumor growth. The protein’s modulation of AMPK-mTOR signaling can impact cancer cell survival and response to treatment. Additionally in diabetes Sestrin-2 plays a part in maintaining insulin sensitivity and glucose metabolism often disrupted in diabetic patients. Its ability to regulate oxidative stress and cellular metabolism draws a link between Sestrin-2 and these complex disorders emphasizing its potential as a therapeutic target.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human SESN2 (Sestrin-2) knockout HeLa cell lysate

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Sanger Sequencing - Human SESN2 (Sestrin-2) knockout HeLa cell lysate (AB257665)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Complementary Products

Primary Antibodies

Cell Lines & Lysates

Cell Lines & Lysates

Cell Lines & Lysates

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Zygosity

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise