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AB305280

Human IFIH1 knockout Jurkat cell line

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IFIH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Homozygote, 46 bp deletion. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
2 Images
Western blot - Human IFIH1 knockout Jurkat cell line (AB305280)
  • WB

Lab

Western blot - Human IFIH1 knockout Jurkat cell line (AB305280)

All lanes:

Western blot - Anti-MDA5 antibody [EPR6743] (<a href='/en-us/products/primary-antibodies/mda5-antibody-epr6743-ab126630'>ab126630</a>) at 1/1000 dilution

Lane 1:

Wild-type Jurkat, LPS (1 ug/mL, 24 h) cell lysate at 20 µg

Lane 2:

Wild-type Jurkat, vehicle control LPS (0 ug/mL, 24 h) cell lysate at 20 µg

Lanes 3 - 4:

Western blot - Human IFIH1 knockout Jurkat cell line (ab305280)

Lane 3:

IFIH1 knockout Jurkat, LPS (1 ug/mL, 24 h) cell lysate at 20 µg

Lane 4:

IFIH1 knockout Jurkat, vehicle control LPS (0 ug/mL, 24 h) cell lysate at 20 µg

Predicted band size: 117 kDa,90 kDa

Observed band size: 140 kDa

false

Sanger Sequencing - Human IFIH1 knockout Jurkat cell line (AB305280)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human IFIH1 knockout Jurkat cell line (AB305280)

Homozygote, 46 bp deletion

Key facts

Cell type

Jurkat

Species or organism

Human

Tissue

Blood

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Homozygote, 46 bp deletion

Disease

Non-Hodgkin Lymphoma

Reactivity data

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

Product details

Recommended control: Human wild-type Jurkat cell line (ab275468). 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.

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

Culture medium:  RPMI + 10% FBS

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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 2x105 cells/mL is recommended.
• Do not allow the cell density to exceed 3x106 cells/mL

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.

Properties and storage information

Gene name
IFIH1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage duration
A few days
Appropriate short-term storage conditions
-80°C
Appropriate long-term storage conditions
-196°C

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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 2x105 cells/mL is recommended.
  • Do not allow cell density to exceed 3x106 cells/mL.
Culture medium

RPMI + 10% FBS

Cryopreservation medium

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

Supplementary information

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

MDA5 also known as melanoma differentiation-associated protein 5 or IFIH1 is a cytosolic sensor involved in the detection of viral RNA. It is a member of the RIG-I-like receptor family and possesses a molecular weight of approximately 116 kilodaltons. MDA5 protein is expressed across various tissues but shows higher expression in the immune system's cells including macrophages and dendritic cells. MDA5 recognizes long double-stranded RNA a common viral replication byproduct triggering immune responses.
Biological function summary

In cellular defense mechanisms MDA5 plays an important role in initiating antiviral responses. It operates as part of a signaling complex that includes MAVS (mitochondrial antiviral-signaling protein) leading to the production of type I interferons and pro-inflammatory cytokines. These responses promote an antiviral state in host cells aiding in the containment and clearance of viral infections. MDA5 antibodies can be useful tools for studying these immune processes.

Pathways

MDA5 interacts with antiviral signaling pathways such as the IFN signaling pathway and the RIG-I-like receptor signaling pathway. Upon recognizing viral RNA MDA-5 cooperates with proteins like RIG-I and MAVS to activate downstream signaling cascades that mediate immune responses. This collaboration enables efficient viral recognition and response limiting viral replication and spread.

MDA5 has associations with autoimmune diseases like systemic lupus erythematosus and certain types of dermatomyositis. These conditions often involve autoantibodies against MDA5 indicating a possible role in pathogenesis. In these diseases altered function or regulation of related proteins including IFN-induced proteins may contribute to chronic inflammation and autoimmunity. Understanding MDA5's involvement in these contexts is essential for developing therapeutic strategies.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

Product protocols

Product promise

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