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AB265978

Human SEC16A (SEC16) knockout HeLa cell line

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SEC16A 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 SEC16A (SEC16) knockout HeLa cell line (AB265978)
  • Sanger seq

Unknown

Sanger Sequencing - Human SEC16A (SEC16) knockout HeLa cell line (AB265978)

Allele-2 : 1 bp insertion in exon 3.

Sanger Sequencing - Human SEC16A (SEC16) knockout HeLa cell line (AB265978)
  • Sanger seq

Unknown

Sanger Sequencing - Human SEC16A (SEC16) knockout HeLa cell line (AB265978)

Allele-1 : 1 bp deletion in exon 3.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

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

Adenocarcinoma

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
SEC16A
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.

SEC16A also known as SEC16 is an important component in the secretory pathway specifically in vesicle trafficking from the endoplasmic reticulum (ER) to the Golgi apparatus. This protein plays a significant role in the assembly of coat protein complex II (COPII)-coated vesicles acting as a scaffold for other components. SEC16A has a molecular mass of approximately 240 kDa. It is broadly expressed in various tissues with notable expression in cells with high secretory activity.
Biological function summary

Functions of SEC16A are critical in maintaining ER exit sites (ERES) where the budding of COPII vesicles occurs. It localizes to these ERES facilitating the formation of transport vesicles. SEC16A does not work alone; it interacts with several other proteins to form a functional multi-protein complex associated with this vesicle trafficking process. These interactions ensure the efficient transport of proteins and lipids necessary for cellular homeostasis and secretion.

Pathways

SEC16A integrates into the COPII-mediated vesicle formation pathway pivotal for protein sorting and transportation. This protein collaborates closely with the SAR1 SEC23 SEC24 SEC13 and SEC31 proteins through the vesicle formation cycle. Besides its role in COPII assembly SEC16A also contributes to the regulation of ER-Golgi transport influencing the growth factor signaling pathways that depend on the accurate delivery of their receptor components to the cell surface.

Disruptions in the function of SEC16A links to conditions like Anderson-Fabry disease and congenital disorders of glycosylation. Abnormal vesicle trafficking due to SEC16A impairment can lead to an accumulation of metabolites or misfolded proteins that disrupt cellular function. SEC16A's relationship with proteins involved in lipid metabolism and glycosylation supports the role in these disorders. Understanding the behaviour of SEC16A in these contexts may contribute to targeted therapeutic strategies.

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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