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AB266183

Human PIGT knockout HEK-293T cell line

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PIGT KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 5 bp deletion in exon 1.

View Alternative Names

CGI 06, FLJ41596, GPI transamidase component PIG T, GPI transamidase subunit, MGC8909, NDAP, Neurotrophin regulated neuronal development associated protein, Phosphatidyl inositol glycan class T, Phosphatidylinositol glycan anchor biosynthesis class T

2 Images
Sanger Sequencing - Human PIGT knockout HEK-293T cell line (AB266183)
  • Sanger seq

Unknown

Sanger Sequencing - Human PIGT knockout HEK-293T cell line (AB266183)

Allele-1 : 5 bp deletion in exon 1

Sanger Sequencing - Human PIGT knockout HEK-293T cell line (AB266183)
  • Sanger seq

Unknown

Sanger Sequencing - Human PIGT knockout HEK-293T cell line (AB266183)

Allele-2 : 1 bp deletion in exon 1.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 5 bp deletion in exon 1

Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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.

What's included?

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Properties and storage information

Gene name
PIGT
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.
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.

Supplementary information

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

PIGT (Phosphatidylinositol Glycan Anchor Biosynthesis Class T) plays a mechanical role in the biosynthesis of glycosylphosphatidylinositol (GPI) anchors. Also known as PIG-T this protein is part of the GPI transamidase complex and has a molecular mass of approximately 68 kDa. It resides in the endoplasmic reticulum where it supports the attachment of various proteins to the cell membrane through GPI anchoring. PIGT is ubiquitously expressed across different human tissues indicating its fundamental role in cellular physiology.
Biological function summary

PIGT acts as a subunit within the GPI transamidase complex that enhances the transfer of pre-assembled GPI units to proteins. This activity anchors proteins to the cell surface which is necessary for their stability and function. The GPI anchor allows effective signaling adhesion and protection of proteins impacting many cellular processes. The interdependency with other subunits such as PIGA PIGK and PIGS ensures the complete function of the GPI transamidase complex.

Pathways

PIGT remains key within the GPI-anchor biosynthesis pathway playing a critical role in cell surface protein localization. It interacts with other members like PIGU and GAA1 within this pathway forming interactions necessary for proper transit and attachment of GPI-anchored proteins. The functionality of the GPI-anchor pathway directly influences cellular communication through pathways like signal transduction which are essential for different biological processes.

Mutations or deficiencies in PIGT link to Paroxysmal Nocturnal Hemoglobinuria (PNH) and Early Infantile Epileptic Encephalopathy (EIEE). PNH involves defective GPI-anchored proteins affecting the regulation of cell membrane stability while EIEE relates to neurological complications due to disturbed protein functioning. PIGT connects with other proteins like PIGM and PIGL contributing to the disorders' pathophysiology when disruptions in GPI-anchor biosynthesis occur.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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