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AB265669

Human SESN2 (Sestrin-2) knockout HeLa cell line

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SESN2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 4.

View Alternative Names

DKFZp761M0212, DKFZp761M02121, Hi95, Hypoxia induced gene 95, MGC11758, OTTHUMP00000003641, OTTMUSP00000010184, RP23 63D8.3, SES2, SESN2_HUMAN, SEST2, Sestrin-2

3 Images
Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell line (AB265669)
  • WB

Lab

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell line (AB265669)

Lanes 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 2 in wild-type HeLa cells. Loss of signal was observed when knockout sample ab257665 was used. Wild-type and 2 knockout samples were subjected to SDS-PAGE. ab178518 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 (ab265669)

Lane 3:

HeLa 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: 54 kDa

Observed band size: 54 kDa

false

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell line (AB265669)
  • WB

Lab

Western blot - Human SESN2 (Sestrin-2) knockout HeLa cell line (AB265669)

Lanes 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.

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 (ab265669)

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

false

Sanger Sequencing - Human SESN2 (Sestrin-2) knockout HeLa cell line (AB265669)
  • Sanger seq

Unknown

Sanger Sequencing - Human SESN2 (Sestrin-2) knockout HeLa cell line (AB265669)

Homozygous : 1 bp deletion in exon 4.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 4

Disease

Adenocarcinoma

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Reactivity data

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Product details

Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

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
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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

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Target data

See full target information SESN2
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Complementary Products

Primary Antibodies

AB178518

Anti-SESN2/Sestrin-2 antibody [EPR18907]

RabMAb|Recombinant|KO Validated

Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] (AB178518)

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Cell Lines & Lysates

AB265054

Human SIRT6 knockout HeLa cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human SIRT6 knockout HeLa cell line (AB265054)

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Cell Lines & Lysates

AB269650

Human SESN2 (Sestrin-2) knockout HEK-293 cell lysate

Western blot - Human SESN2 (Sestrin-2) knockout HEK-293 cell lysate (AB269650)

  • 0
  • 0
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20

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