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AB264680

Human VCPIP1 (VCIP 135) knockout HeLa cell line

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VCPIP1 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.

View Alternative Names

DUBA3, Deubiquitinating protein VCIP135, KIAA1850, VCIP135, VCIP1_HUMAN, VCP/p47 complex-interacting 135-kDa protein, Valosin containing protein (p97)/p47 complex interacting protein 1, Valosin-containing protein p97/p47 complex-interacting protein 1, Valosin-containing protein p97/p47 complex-interacting protein p135, Vcpip1

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Sanger Sequencing - Human VCPIP1 (VCIP 135) knockout HeLa cell line (AB264680)
  • Sanger seq

Unknown

Sanger Sequencing - Human VCPIP1 (VCIP 135) knockout HeLa cell line (AB264680)

Allele-1 : 1 bp insertion in exon 1.

Sanger Sequencing - Human VCPIP1 (VCIP 135) knockout HeLa cell line (AB264680)
  • Sanger seq

Unknown

Sanger Sequencing - Human VCPIP1 (VCIP 135) knockout HeLa cell line (AB264680)

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

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 insertion in exon 1 and Insertion of the selection cassette in exon 1

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255448). 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, 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
VCPIP1
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.

VCIP135 also known as VCP-interacting protein p135 plays an important role in the process of membrane fusion. This protein has a mass of approximately 135 kDa and is found mainly in the cytosol and nucleus of cells. It directly interacts with valosin-containing protein (VCP) which is important for various cellular functions. The expression of VCIP135 has been noted across multiple tissues highlighting its diverse involvement in cellular activities.
Biological function summary

VCIP135 facilitates endoplasmic reticulum and Golgi membrane fusion by interacting within the VCP complex. In this mechanism it regulates the dynamics of vesicle trafficking and processing influencing the maintenance of cellular homeostasis. By ensuring proper fusion and reassembly of vesicular membranes VCIP135 impacts how well a cell manages its transport and secretory pathways. Its function in coordinating membrane systems confirms its integral role in cell integrity.

Pathways

VCIP135 significantly contributes to the ER-associated degradation (ERAD) and protein ubiquitination pathways. During these processes VCIP135 collaborates closely with proteins such as ubiquitin and VCP to direct the proper destruction of misfolded proteins. This cooperation ensures that the cell adequately manages protein quality control preventing the build-up of harmful nonfunctional proteins. These processes are essential in safeguarding cellular health and consistent workflow of molecular machinery.

Improper function of VCIP135 links to neurodegenerative disorders such as Parkinson's disease and inclusion body myopathy. These conditions often involve protein aggregation due to deficient ubiquitin-proteasome system activity a pathway in which VCIP135 retains functionality through its interaction with VCP. Aberrations in this pathway lead to toxic protein build-up which in turn impacts neuron and muscle cell survival. Understanding VCIP135's role in these diseases aids in developing therapeutic strategies toward modulation of its activity.

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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