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AB275834

Human PRAME knockout U-2 OS cell line

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PRAME KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 100%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

4930534P07Rik, CT130, Cancer/testis antigen 130, MAPE, Melanoma antigen preferentially expressed in tumors, OIP-4, OPA-interacting protein 4, Opa interacting protein OIP4, PRAME_HUMAN, Preferentially expressed antigen in melanoma, Preferentially expressed antigen of melanoma, RP23-250F8.3

1 Images
Next Generation Sequencing - Human PRAME knockout U-2 OS cell line (AB275834)
  • NGS

Supplier Data

Next Generation Sequencing - Human PRAME knockout U-2 OS cell line (AB275834)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 100%

Key facts

Cell type

U-2 OS

Species or organism

Human

Tissue

Bone

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 100%

Disease

Osteosarcoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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
PRAME
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
-196°C|-80°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

McCoY5a + 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.

PRAME also known as Preferentially Expressed Antigen in Melanoma is a protein typically weighing around 50 kDa. It is found mostly in testis tissue but also appears in several types of tumors including melanoma and some leukemias. PRAME does not show significant expression in most normal tissues making it a potential target for cancer therapies. Immunohistochemistry (IHC) techniques such as PRAME IHC and PRAME staining often use monoclonal antibodies like mAb EPR20330 to detect PRAME protein presence in various samples.
Biological function summary

PRAME acts as a repressor of retinoic acid signaling a process important for cell differentiation and growth. It does not directly form part of a complex but interacts with components involved in retinoic acid pathways. By binding to retinoic acid receptor complexes PRAME prevents activation of genes involved in cell cycle arrest and apoptosis contributing to unchecked cellular proliferation seen in certain cancers.

Pathways

PRAME participates in the retinoic acid and various oncogenic signaling processes. It influences the retinoic acid pathway by interfering with the retinoic acid receptor (RAR) signaling. This interference with key proteins like RAR disrupts the normal regulatory processes that typically inhibit cancer progression. PRAME's modulation of these pathways highlights its role in promoting tumor growth and survival.

PRAME has significant relevance to cancer particularly melanoma. It is often used as a biomarker for melanoma identification due to its overexpression in such tumors. Additionally PRAME's connection to acute myeloid leukemia (AML) emerges from its ability to act similarly in diverse malignant cells. In these diseases the overexpression of PRAME interferes with normal differentiation processes and allows for sustained proliferation of malignant cells suggesting its potential role as a therapeutic target or diagnostic marker in oncology.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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