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AB264013

Human IRS2 knockout HEK-293 cell line

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IRS2 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Immunocytochemistry/ Immunofluorescence - Human IRS2 knockout HEK-293 cell line (AB264013)
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Immunocytochemistry/ Immunofluorescence - Human IRS2 knockout HEK-293 cell line (AB264013)

ab134101 staining IRS2 in wild-type HEK-293 cells (top panel) and IRS2 knockout HEK-293 cells (ab264013) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab134101 at 1/500 and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Western blot - Human IRS2 knockout HEK-293 cell line (AB264013)
  • WB

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Western blot - Human IRS2 knockout HEK-293 cell line (AB264013)

Lanes 1 - 4 : Merged signal (red and green). Green - ab134101 observed at 160 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab134101 was shown to react with IRS2 in wild-type HEK-293 cells in western blot with loss of signal observed in IRS2 knockout sample. Wild-type and IRS2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134101 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IRS2 antibody [EPR904(2)] (<a href='/en-us/products/primary-antibodies/irs2-antibody-epr9042-ab134101'>ab134101</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

IRS2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human IRS2 knockout HEK-293 cell line (ab264013)

Lane 3:

A-375 cell lysate at 20 µg

Lane 4:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 137 kDa,25 kDa

Observed band size: 160 kDa

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Western blot - Human IRS2 knockout HEK-293 cell line (AB264013)
  • WB

Lab

Western blot - Human IRS2 knockout HEK-293 cell line (AB264013)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133276 observed at 160 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab133276 was shown to react with IRS2 in wild-type HEK-293 cells in western blot with loss of signal observed in IRS2 knockout sample. Wild-type and IRS2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab133276 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IRS2 antibody [EPR1650(2)] (<a href='/en-us/products/primary-antibodies/irs2-antibody-epr16502-ab133276'>ab133276</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

IRS2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human IRS2 knockout HEK-293 cell line (ab264013)

Lane 3:

A-375 cell lysate at 20 µg

Lane 4:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 137 kDa

Observed band size: 160 kDa

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Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Immunocytochemistry,Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9

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Complementary Products

Primary Antibodies

AB134101

Anti-IRS2 antibody [EPR904(2)]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (AB134101)

  • 0
  • 0
Cell Lines & Lysates

AB264016

Human LPL (Lipoprotein lipase) knockout HeLa cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human LPL (Lipoprotein lipase) knockout HeLa cell line (AB264016)

  • 0
  • 0
Primary Antibodies

AB231441

Anti-IL-2 Receptor alpha antibody [SP176]

20ul selling size|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-2 Receptor alpha antibody [SP176] (AB231441)

  • 3
  • 5
Primary Antibodies

AB189524

Anti-CD44 antibody [EPR18668]

RabMAb|Recombinant|Lab Essentials|20ul selling size|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (AB189524)

  • 5
  • 1
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Reactivity data

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

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
IRS2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Next Generation Sequencing, Western blot
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.

Insulin Receptor Substrate 2 (IRS2) is an important cytoplasmic adaptor protein involved in insulin signaling. Alternate names for IRS2 include IRS-2 or HEK293 products IRS2. It plays a role in transducing signals from the insulin receptor tyrosine kinase to downstream effectors. IRS2 has a molecular mass of approximately 180 kDa. Expression of IRS2 occurs in many tissues with significant presence in liver and skeletal muscle indicating its broad physiological role.
Biological function summary

IRS2 acts in signal transduction and is a part of the insulin and insulin-like growth factor signaling pathways. IRS2 does not function alone but interacts with other molecules to propagate signals that regulate cellular processes. It is essential for glucose homeostasis and lipid metabolism playing a critical role in maintaining energy balance within the organism. IRS2 also mediates other biological processes like cell growth and differentiation.

Pathways

With regards to pathways IRS2 is involved in the insulin signaling and mTOR pathways. It serves as an important junction between metabolic and growth-promoting signals. Activation of IRS2 through insulin receptors leads to downstream signaling involving PI3K and Akt essential for mediating metabolic actions of insulin. It also interacts with mTOR a central protein in cell growth regulation. These pathways illustrate the integration of IRS2 in broader cellular responses to growth signals and energy availability.

IRS2 is linked to conditions like type 2 diabetes and obesity. Dysregulation of IRS2-mediated signaling can contribute to insulin resistance a characteristic feature of type 2 diabetes. In such disorders IRS2 signaling may be impaired or altered affecting its interaction with proteins like PI3K and Akt which are also implicated in these conditions. Furthermore chronic imbalance in IRS2 activity may contribute to obesity as it affects key metabolic pathways that regulate fat storage and energy expenditure.
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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 Insulin receptor substrate 2
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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