DU145 cell line was obtained from the American Tissue Type Collection (ATCC, Manassas, VA). The cell line was cultured in Iscove’s modified Dulbecco’s medium (IMDM) from Mediatech (Manassas,VA) supplemented with 10% fetal bovine serum (FBS) Hyclone Laboratories (Novato, CA) and incubated at 37° in a 5% CO2 humidified chamber. Non-enzymatic Cellstripper (CelGro, Manassas, VA) was used for cell harvesting. Antibodies used for immunofluorescence microscopy include: anti-α6 integrin (A6NT, (21 (link))), anti-Desmin (Atlas Antibodies HPA018803), anti-E-Cadherin M168 (Abcam ab76055), anti-CK5/14 (KA2), anti-CK18 (Abcam EPR1626), and anti-FOXA1 (Abcam EPR10881). Antibodies used for flow cytometry include: anti-α6 integrin phycoerythrin (PE) conjugated GoH3 (eBioscience, San Diego, CA) and ADG3937 (Sekisui Diagnostics) mouse monoclonal antibody against uPAR. Alexa Fluor 488 (Invitrogen) secondary antibody was used for uPAR detection.
Adg3937
Manufactured by Sekisui
Sourced in Germany
The ADG3937 is a laboratory equipment product designed for general laboratory applications. It serves as a core function for its intended purpose.
Automatically generated - may contain errors
Lab products found in correlation
Epr10881, by Abcam (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Peroxidase block, by Agilent Technologies (2 mentions)
K4003, by Agilent Technologies (2 mentions)
K4001, by Agilent Technologies (2 mentions)
S2001, by Agilent Technologies (2 mentions)
K3469, by Agilent Technologies (2 mentions)
S2020, by Agilent Technologies (2 mentions)
Target retrieval solution, by Agilent Technologies (2 mentions)
2 protocols using adg3937
1
DU145 Cell Line Characterization
2
Immunohistochemical Analysis of uPAR in Melanoma
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The immunohistochemical staining was performed on 5 μm TMA sections of paraffin-embedded archival tissue. Sufficient tumor tissue for immunohistochemistry was available in 248 of the primary melanoma cases and 68 of the loco-regional metastatic melanomas. The slides were dewaxed with xylene/ethanol. Antigen retrieval was performed for 20 minutes in Target Retrieval Solution (DAKO 1699) (pH = 6) in microwave. Endogenous peroxidase activity was prevented by treating the slides with peroxidase block (DAKO S2001) for 8 minutes. The slides were incubated with the mouse monoclonal antibody uPAR (dilution 1:100) (ADG 3937, Sekisui Diagnostics, Pfungstadt, Germany) overnight at 4°C. EnVision labelled polymer method was then used (DAKO K4001 or K4003) for 30 minutes. 3-amino-9-ethylcarbazole (AEC) (DAKO K3469) was used as substrate chromogen. Brief counterstaining was performed with hematoxylin (DAKO S2020). Negative control was obtained by omitting the primary antibody.
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