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AB267190

Human GBP3 knockout A549 cell line

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GBP3 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3. 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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Sanger Sequencing - Human GBP3 knockout A549 cell line (AB267190)
  • Sanger seq

Unknown

Sanger Sequencing - Human GBP3 knockout A549 cell line (AB267190)

Allele-1 : 1 bp insertion in exon3

Sanger Sequencing - Human GBP3 knockout A549 cell line (AB267190)
  • Sanger seq

Unknown

Sanger Sequencing - Human GBP3 knockout A549 cell line (AB267190)

Allele-2 : 1 bp deletion in exon 3.

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3

Disease

Carcinoma

Product details

We will provide viable cells that proliferate on revival.

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

What's included?

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Properties and storage information

Gene name
GBP3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°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 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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 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.

The protein known as GBP3 also called Guanylate Binding Protein 3 is a member of the guanylate-binding protein family. This protein has a molecular mass of approximately 67 kDa. It mainly functions in cellular processes involving GTP hydrolysis which plays a significant role in certain immune responses. GBP3 expresses in various tissues including the heart placenta and lungs showing higher presence in immune-related cells and tissues.
Biological function summary

GBP3 contributes to cellular defense by promoting anti-viral states and mediating immune signaling. It has been found to associate with large protein complexes that enhance its stability and activity during inflammation. This involvement supports various immune functions and contributes to the body's response to pathogenic threats. Such actions highlight GBP3's importance in maintaining cellular immunity and promoting pathogenic resistance.

Pathways

GBP3 interacts within the interferon-gamma signaling pathway where it facilitates host defense mechanisms. The protein's activity aligns closely with the JAK-STAT pathway important for the transduction of extracellular signals from the environment to the cell nucleus. GBP3 operates alongside other proteins such as GBP1 and GBP5 which are also part of guanylate-binding proteins involved in immune regulation and protection against intracellular bacteria and viruses.

Impairments in GBP3 function have connections to infectious diseases and inflammatory disorders. Studies suggest that altered GBP3 expression might relate to an increased susceptibility to viral infections such as hepatitis. Additionally abnormalities in this protein may associate with chronic inflammatory conditions like Crohn's disease. GBP3 interacts with proteins like STAT1 and GBP1 which play critical roles in modulating immune response and maintaining homeostasis within inflammatory conditions.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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