WM266-4 Viable Cells - WM266-4-01-0001

Referentie WM266-4-01-0001

Formaat : 1mL

Merk : Rockland Immunochemicals

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Specifications for WM266-4 Viable Cells

Product Details

WM266-4 Viable Cells - WM266-4-01-0001
Melanoma cells, patient derived tumor cells, tumor models, skin cancer testing, xenograft production, Human melanoma tumor cell
Human

Target Details

Cells are sterile, validated by short tandem repeat profiling, and are tested as negative for mycoplasma. It is recommended that cell lines are tested for mycoplasma contamination and short tandem repeat (STR) profiling every 10 passages or each time a frozen seed stock is made. See cell culture protocol for additional details.

Application Details

Cellular Assay, ELISA, IF, IHC, Other, WB  - View References
The key applications of these cell lines include genetic studies, xenograft production, drug testing, and drug target discovery. These cell line models can be used in various biological assays, and for identifying critical target genes, and cell signaling pathways.

Cell Line Data

Human Melanoma
Viable Cells
small flat mesenchymal
V600D
WT
WT
Hemizygous Deletion
Tumor Specialized Media with 2% HI-FBS
Cells should be maintained between 30 – 95% confluence in tumor specialized medium with 2% FBS; split cultures 1:3 every 7 days using 0.25% trypsin/EDTA.
36°C with 5% CO2

Formulation

1.0 million cells/mL Count By Hemocytometer
None
None
None

Shipping & Handling

Dry Ice
Cells are frozen with 90% FBS/10% DMSO solution at about 1x10^6 cells/ml. Store vial in liquid nitrogen upon arrival.
Expiration date is two (2) years from date of receipt.

Background

WM266-4 is a metastatic human melanoma cell line with small flat mesenchymal morphology. This cell line was derived from the same patient as the cell lines WM115, WM239A, and WM165-1. WM115 cell line originated from the primary tumor, and WM266-4, WM165-1 and WM239A were from individual lymph-node metastases. The subject (55-year-old female) displayed VGP with a Clark level III tumor with thickness of 2.24mm.This cell line features the specific V600D (Val600Asp) mutation at codon 600 in the BRAF gene. This cell line also expresses PTEN loss of function including hemizygous deletion. WM266-4 cell line is wild type for N-RAS, c-KIT, and CDK4 genes. WM266-4 cells produce xenograft tumors when injected into immunocompromised mice.

References (5)

(). Cytokine and Chemokine Receptor Patterns of Human Malignant Melanoma Cell Lines. Int J Mol Sci.
Applications
RNA seq, qPCR, Reverse Transcription, RT-PCR Analysis; Statistical Analysis; In Vitro Assay
PubMed
(). ---Inhibiting FAK-Paxillin Interaction Reduces Migration and Invadopodia-Mediated Matrix Degradation in Metastatic Melanoma Cells. Cancers (Basel).
Applications
IF, Confocal Microscopy; WB, IB, PCA
PubMed
(). Epigenetic silencing of CDR1as drives IGF2BP3-mediated melanoma invasion and metastasis. Cancer Cell.
Applications
Other
PubMed
(). Melanoma Progression Inhibits Pluripotency and Differentiation of Melanoma-Derived iPSCs Produces Cells with Neural-like Mixed Dysplastic Phenotype. Stem Cell Reports.
Applications
Cell Proliferation Assay
PubMed
(). Targeting the Proteasome-Associated Deubiquitinating Enzyme USP14 Impairs Melanoma Cell Survival and Overcomes Resistance to MAPK-Targeting Therapies. Mol Cancer Ther.
Applications
EIA, ELISA
PubMed
Frequently Asked Questions (8)

How can I ensure that the cells will be viable after resuspension?

Complete adherence to the protocol is necessary to ensure the viability of the cells. Cells should be maintained between 30-95% confluence in Tumor Specialized media with 2% FBS (heat inactivated) unless otherwise noted in cell line specific instructions.

How should I handle the cells before culturing?

Before thawing, frozen cells can be stored in liquid nitrogen or at -80°C only. To thaw the melanoma cells, move the vial from liquid nitrogen to dry ice immediately. Melanoma cells should be thawed quickly. Freeze thaw cycles can damage / kill the cells and should be avoided. Centrifuge the cells at 1500 rpm (500 x g) for 5 minutes at room temperature. Discard the supernatant. Resuspend cell pellet in 5 mL of Tumor Specialized Media with FBS (heat inactivated) and transfer to a T-25 flask. Place the flask in a humidified incubator (5% CO2) at 36°C overnight. Check the flask after 24 hours for attachment of cells to the flask. If cells are attached, remove the media and add fresh media to the flask.

Which media should I use to grow melanoma cells?

Tumor specialized Media with 2% FBS is preferred media for melanoma cell lines. However, alternative media (not preferred) recommendations include: 1. DMEM with 5% FBS; 2. RPMI with 5% FBS.

Do you have recommended sources for Tumor Specialized Media components?

The recommended manufacturer for each component of the Tumor Specialized Media are: MCDB-153 (Sigma-Aldrich, item# M-7403), Leibovitz’s L-15 (Sigma-Aldrich, item# L1518), FBS (heat inactivated) (Rockland, item# FBS-010-0100), Calcium Chloride (Sigma-Aldrich, item# C5670), Sodium Bicarbonate [Added to MCDB-153 preparation] (Sigma-Aldrich, item# S5761). Please consult the protocol for detailed instructions on the preparation of the media.

What is the optimal growth temperature for melanoma cells?

Melanoma cells grow best at 36°C (not 37°C) with 5% CO2. These incubation conditions must be followed exactly to ensure the viability and growth of the cells.

What is the recommended media for freezing viable melanoma cells?

Freezing media containing 90% FBS and 10% DMSO. Thawed cells should not be frozen again. Melanoma cells should be frozen slowly but thawed quickly.

What is the source of the melanoma cell lines?

The melanoma cell lines were established from patient-derived tumors. Information about the patients or treatments received is not available.

Are melanoma cell lines immortalized?

Melanoma cell lines are considered immortalized because they can replicate indefinitely.

Related Protocols

Adherent Cell Lysis Protocol
Azide Removal by Dialysis Protocol
ELISA Protocol
Fluorescent Western Blotting Protocol
Heat-induced Antigen Retrieval Protocol
Histone Immunoblotting Protocol
Immunocytochemistry (ICC) Protocol
Immunofluorescence (IF) Protocol
Immunohistochemistry (IHC) Protocol
In-Cell Western (ICW) Protocol
IP-WB with TrueBlot® Protocol
Multi-Lysate Western Blotting Protocol
Nuclear & Cytoplasmic Extract Protocol
Protease-induced Antigen Retrieval Protocol
Sandwich ELISA Protocol for Collagen
Staining Paraffin Sections by PAP Procedure
Suspension Cultured Cell Lysis Protocol
Western Blotting (WB) Protocol

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Disclaimer

No test method can provide total assurance that the hepatitis B virus, hepatitis C virus, human immunodeficiency virus, or any other infectious agents are absent. Thus, all blood products, including purified proteins derived from human blood sources, should be handled at Biosafety Level 2 as recommended by the CDC\NIH manual entitled Biosafety in Microbiological and Biomedical Laboratories for potentially infectious human serum, blood specimens or proteins derived from same. Source material for the human blood product supplied to your facility has been tested for the detection of HIV antibody, Hepatitis B surface antigen, antibody to Hepatitis C, HIV 1 antigen(s), antibody to HTLV - I/II, and syphilis by FDA guidelines. All units were found to be non-reactive/negative for these tests. All human blood source material is collected in FDA licensed centers and is tested with FDA approved test kits.; Cell Line Limited Use License Required. THIS PRODUCT IS SUBJECT TO AN END-USER LICENSE AGREEMENT (EULA). BY ACCEPTING THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF USE SET FORTH BELOW and SET FORTH IN THE EULA. THIS PRODUCT IS FOR IN VITRO RESEARCH USE ONLY. THERAPEUTIC, DIAGNOSTIC, OR VETERINARY USE IS PROHIBITED. This product may not be resold or transferred by the recipient and may be used only by the recipient, in the recipient’s facility and only for research use and other uses specifically permitted by the EULA. No other commercial use is allowed. “Commercial Use” means any and all uses of this product by recipient or others for monetary or other consideration, including providing services, supplying information or data to unaffiliated third parties, and resale or transfer of this product for any use. Recipient has no right to modify, derivatize, genetically engineer or otherwise create variations of this product or associated cells or cell lines. ROCKLAND AND WISTAR MAKE NO REPRESENTATIONS AND EXTEND NO WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR THAT THE USE OF THE PRODUCTS WILL NOT INFRINGE ANY PATENT, COPYRIGHT, TRADEMARK, OR OTHER PROPRIETARY RIGHTS. The terms set forth herein and in the EULA shall be governed by the laws of the Commonwealth of Pennsylvania, USA. To obtain a COMMERCIAL USE license for this product, please contact Rockland Immunochemicals, Inc. Please contact a technical service representative for more information. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.

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