Wm983a

Manufactured by Rockland Immunochemicals

Sourced in Ireland

The WM983A is a laboratory equipment product manufactured by Rockland Immunochemicals. It is designed to perform specific functions within the laboratory setting. The core function of this product is to assist in the analysis and processing of samples, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

7 protocols using wm983a

Cell Line Maintenance and Validation

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HEK293, A375, WM164 cell lines were purchased from ATCC. WM983A and WM983B cell lines were purchased from Rockland (Philadelphia, PA). Melanocytes (CDKN2A null) were provided by Dr. Ian Robert Watson (McGill University). All cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO) with 4.5 g/liter glucose supplemented with 10% fetal bovine serum (FBS), 100 IU/ml penicillin, and 100 µg/ml streptomycin, under 37 °C/5%CO₂ conditions. Colo829 cells were purchased from ATCC and maintained in Roswell Park Memorial Institute Medium (RPMI) with 10% FBS, 100 IU/ml penicillin and 100 µg/ml streptomycin, under 37 °C/5%CO₂ conditions. Cells were regularly tested by PCR to exclude mycoplasma contamination.

Houles T., Lavoie G., Nourreddine S., Cheung W., Vaillancourt-Jean É., Guérin C.M., Bouttier M., Grondin B., Lin S., Saba-El-Leil M.K., Angers S., Meloche S, & Roux P.P. (2022). CDK12 is hyperactivated and a synthetic-lethal target in BRAF-mutated melanoma. Nature Communications, 13, 6457.

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Cell Line Maintenance and Authentication

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HEK293, A375, WM164 cell lines were purchased from ATCC. WM983A and WM983B cell lines were purchased from Rockland (Philadelphia, PA). All cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO) with 4.5 g/liter glucose supplemented with 10% fetal bovine serum (FBS), 100 IU/mL penicillin, and 100 µg/mL streptomycin, under 37 °C/5%CO₂ conditions. Colo829 cells were purchased from ATCC and maintained in Roswell Park Memorial Institute Medium (RPMI) with 10% FBS, 100 IU/mL penicillin and 100 µg/mL streptomycin, under 37 °C/5%CO₂ conditions. Cells were regularly tested by PCR to exclude mycoplasma contamination.

Houles T., Boucher J., Lavoie G., MacLeod G., Lin S., Angers S, & Roux P.P. (2023). The CDK12 inhibitor SR-4835 functions as a molecular glue that promotes cyclin K degradation in melanoma. Cell Death Discovery, 9, 459.

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Melanoma Cell Line Authentication and Culture

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The following cell lines were from the American Tissue Type Collection: A375, HEMN (normal adult epidermal melanocytes) and Jurkat. The following cell lines were from Rockland Immunochemicals: WM983A, WM983B, WM1366, WM115, WM266-4, WM164, WM793 and Lu1205. SW1 melanoma cells (gift from the Ronai laboratory at the Sanford Burnham Prebys Medical Discovery Institute) and SM1 melanoma cells (gift from the Smalley laboratory at the Moffitt Cancer Center) were cultured in DMEM containing 10% FBS, 1 g ml⁻¹ glucose and 4 mM l-glutamine at 37 °C with 5% CO₂. HEMN cells were grown in Lonza MGM-4 growth medium; before collection for IB analysis, the cells were switched to the same medium as the other cells overnight. Cell lines were transfected using Lipofectamine 2000 (Invitrogen). Primary CD4⁺ T cells were collected using the EasySep Human CD4⁺ negative selection isolation kit (Stemcell Technologies) according to the manufacturer’s protocols. Upon arrival at the laboratory, all cell lines are quarantined until they have passed footprint identification and mycoplasma testing (as mycoplasma negative). The identities of all cell lines (human and mouse) in the Lau laboratory are verified annually by short tandem repeat-based authentication ‘CellCheck’ services provided through IDEXX BioResearch.

Lester D.K., Burton C., Gardner A., Innamarato P., Kodumudi K., Liu Q., Adhikari E., Ming Q., Williamson D.B., Frederick D.T., Sharova T., White M.G., Markowitz J., Cao B., Nguyen J., Johnson J., Beatty M., Mockabee-Macias A., Mercurio M., Watson G., Chen P.L., McCarthy S., MoranSegura C., Messina J., Thomas K.L., Darville L., Izumi V., Koomen J.M., Pilon-Thomas S.A., Ruffell B., Luca V.C., Haltiwanger R.S., Wang X., Wargo J.A., Boland G.M, & Lau E.K. (2023). Fucosylation of HLA-DRB1 regulates CD4+ T cell-mediated anti-melanoma immunity and enhances immunotherapy efficacy. Nature Cancer, 4(2), 222-239.

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Cell Culture of Melanoma and Keratinocyte Lines

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The human melanoma cells lines A375 were bought from Sigma–Aldrich (Milan, Italy). WM1862, WM983A and WM983B cell lines were purchased from Rockland (Limerick, Ireland). Human keratinocytes (HaCaT) were purchased from Lonza (Basel, Switzerland)). All cell lines were cultured in RPMI 1640 medium with GlutaMAXTM and were supplemented with 10% heat-inactivated fetal calf serum, 2 mmol/L L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin and 10 mM HEPES buffer (all from Gibco; New York, NY, USA). Cells were grown at 37 °C in a humidified incubator under 5% CO₂. All cell lines used in this study were characterized by the cell bank where they were purchased. ERU was purchased from Cayman Chemicals (Michigan, CA, USA).

Maresca D.C., Conte L., Romano B., Ianaro A, & Ercolano G. (2022). Antiproliferative and Proapoptotic Effects of Erucin, A Diet-Derived H2S Donor, on Human Melanoma Cells. Antioxidants, 12(1), 41.

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Cultivation of Melanocytes and Melanoma Cells

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Normal human epidermal melanocytes (NHEM) were purchased from Lonza (Walkersville, MD, USA) and were cultured in melanocyte growth medium 2 (Lonza). The A375 human melanoma cell lines were purchased from Sigma–Aldrich (Milan, Italy) and were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 2 mmol/L L-glutamine, 100 U/mL penicillin, 100 µg/mL streptomycin and 10 mM HEPES buffer (all from Gibco; New York, NY, USA). WM983A and WM983B cell lines were purchased from Rockland (Limerick, Ireland) and cultured in complete RPMI 1640 Medium. Cells were incubated at 37 °C in a humidified incubator under 5% CO2.

Romano B., Maresca D.C., Somma F., Ahmadi P., Putra M.Y., Rahmawati S.I., Chianese G., Formisano C., Ianaro A, & Ercolano G. (2023). Ircinia ramosa Sponge Extract (iSP) Induces Apoptosis in Human Melanoma Cells and Inhibits Melanoma Cell Migration and Invasiveness. Marine Drugs, 21(7), 371.

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Melanoma Cell Lines Characterization

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The human melanoma cell lines A375 cells were purchased from Sigma‐Aldrich (Milan, Italy), WM983A, and WM983B cells were purchased from Rockland (Limerick, Ireland). The murine melanoma cells B16F10 were purchased and characterized from IRCCS AOU San Martino–IST (Genova, Italy). Normal human epidermal melanocytes (NHEMs) were purchased from Lonza (Walkersville, Maryland) and were grown in melanocyte growth medium 2 (Lonza). A375 and B16F10 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum, 2 mmol/L L‐glutamine, 100 μmol/L non‐essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml), and 1 mmol/L sodium pyruvate (all from Sigma‐Aldrich, Milan, Italy). WM983A and WM983B were cultured in Tumor Specialized Media (1:5 Leibovitz's—MCDB153), containing 2% Inactivated FBS and 1.68 mM CaCl₂. Cells were grown at 37°C in a humidified incubator under 5% CO₂. All cell lines used in this study were characterized by the cell bank where they were purchased.

De Cicco P., Ercolano G., Tenore G.C, & Ianaro A. (2022). Olive leaf extract inhibits metastatic melanoma spread through suppression of epithelial to mesenchymal transition. Phytotherapy Research, 36(10), 4002-4013.

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Melanoma Cell Lines Cultivation Protocol

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NHEM were purchased from Lonza (Walkersville, MD, USA) and were grown in Melanocyte growth medium 2 (Lonza). The melanoma cells lines B16/F10, Sk-Mel-5 and Sk-Mel-28 were purchased from IRCCS AOU San Martino – IST (Genova, Italy), A375 from Sigma-Aldrich (Milan, Italy) and were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum, 2 mmol/L L-glutamine, 100 μmol/L non essential amino acids, penicillin (100 U/mL), streptomycin (100 μg/mL) and 1 mmol/L sodium pyruvate (all from Sigma-Aldrich, Milan, Italy). WM35, WM983A and WM983B were from Rockland (Limerick, Ireland) and were cultured in Tumor Specialized Media (1:5 Leibovitz's – MCDB153), containing 2% Inactivated FBS and 1,68 mM CaCl₂. Cells were grown at 37°C in a humidified incubator under 5% CO₂. All cell lines used in this study were characterized by the cell bank were they were purchased. Celecoxib (Selleck Chemicals, Munich, Germany) and naproxen (Sigma-Aldrich, USA) were solubilized in H₂O.

Panza E., De Cicco P., Ercolano G., Armogida C., Scognamiglio G., Anniciello A.M., Botti G., Cirino G, & Ianaro A. (2016). Differential expression of cyclooxygenase-2 in metastatic melanoma affects progression free survival. Oncotarget, 7(35), 57077-57085.

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