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AB266082

Human KNSTRN (SKAP) knockout HEK-293T cell line

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KNSTRN KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp insertion in exon 1 and Insertion of the selection cassette in exon 1. 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 KNSTRN (SKAP) knockout HEK-293T cell line (AB266082)
  • Sanger seq

Unknown

Sanger Sequencing - Human KNSTRN (SKAP) knockout HEK-293T cell line (AB266082)

Allele-2 : Insertion of the selection cassette in exon 1.

Sanger Sequencing - Human KNSTRN (SKAP) knockout HEK-293T cell line (AB266082)
  • Sanger seq

Unknown

Sanger Sequencing - Human KNSTRN (SKAP) knockout HEK-293T cell line (AB266082)

Allele-1 : 4 bp insertion 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, 4 bp insertion in exon 1 and Insertion of the selection cassette in exon 1

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
KNSTRN
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

Supplementary information

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

SKAP also known as Src Kinase-Associated Phosphoprotein of 55 kDa is an adaptor protein that plays an important role in T cell activation. It weighs approximately 55 kDa and is prominently expressed in lymphoid tissues where it supports the immune response. Functionally SKAP forms part of signaling cascades that transmit extracellular signals into the cell interior modulating cellular activities.
Biological function summary

SKAP regulates immune cell function by influencing adhesion and migration processes. It often forms part of larger complexes within the immune synapse facilitating interactions between proteins necessary for efficient signaling. SKAP's role in T cells involves partnering with other signaling molecules to strengthen immune responses against infections by promoting cell adhesion.

Pathways

SKAP participates actively in the T-cell receptor (TCR) signaling and integrin-mediated pathways. It associates with proteins such as LFA-1 an important integrin that mediates cell adhesion. SKAP affects the downstream signaling events by modulating key interactions and conformations which are important for effective immune response initiation and regulation.

SKAP has notable connections with autoimmune diseases like rheumatoid arthritis and multiple sclerosis. Its deregulation affects the immune system's balance leading to exacerbated inflammatory responses. Connections to proteins like RapL which interacts with integrins to regulate cell adhesion and migration imply its involvements in pathological conditions involving the immune system's improper regulation.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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.

Product protocols

Product promise

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