The mouse alveolar macrophage cell line MH-S, and human lung fibroblast MRC-5 cells were obtained from ATCC and cultured in regular DMEM supplemented with 10% FBS (Sigma). For overexpression studies easy-to-transfect human lung osteosarcoma (U2OS) and human embryonic kidney (HEK293) cells were also obtained from ATCC and cultured in DMEM medium with 10% FBS. Human lung adenocarcinoma (NCI-H441 and A549) cells were cultured in RPMI-1640 medium supplemented with 10% FBS. Rabbit polyclonal TTP antibodies were purchased from Abcam Inc. (ab36558 and ab83579). Mouse monoclonal anti-DDK antibody was purchased from Origene (Rockville, MD), and antibodies against β-TrCP1 and GAPDH were purchased from Cell Signaling Technology (Danvers, MA). Control (Cat. No. D-001810) and β-TrCP1 (Cat. No. D-003463) siRNA were purchased from Thermo Fisher Scientific (Lafayette, CO). P38 kinase inhibitor SB203580 (Cat. V1161) was purchased from Promega Corp. (Madison, WI).
Mouse monoclonal anti ddk antibody
Manufactured by OriGene
Sourced in United States
The Mouse monoclonal anti-DDK antibody is a laboratory reagent used to detect and identify proteins tagged with the DDK (FLAG) epitope. It can be used in various immunoassay techniques such as Western blotting, immunoprecipitation, and immunocytochemistry to help analyze protein expression and interactions.
Automatically generated - may contain errors
Lab products found in correlation
Ab83579, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Fibronectin, by R&D Systems (1 mentions)
Recombinant human rhesus macaque feline cxcl12 sdf1α, by R&D Systems (1 mentions)
Mouse monoclonal anti gfp, by Roche (1 mentions)
Mouse monoclonal anti β actin antibody, by Merck Group (1 mentions)
Confocal laser scanning microscopy, by Zeiss (1 mentions)
3 protocols using mouse monoclonal anti ddk antibody
1
Cell Culture Techniques for Overexpression
2
Molecular Characterization of ELMO1
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YFP-tagged ORF clone of ELMO1 was purchased from GeneCopoeia (Rockville, MD). YFP-tagged truncated mutants of ELMO1 were generated by a PCR-based approach and cloned into the Af1II/NotI sites of pReceiver-M15 (GeneCopoeia). The constructs were sequenced to confirm the appropriate mutations. Goat polyclonal anti-ELMO1 and goat polyclonal anti-ELMO2 antibodies were obtained from Abcam (Cambridge, MA). Mouse monoclonal Anti-DDK antibody came from Origene (Rockville, MD). Rabbit polyclonal anti-Gβ (M-14) antibodies and goat polyclonal anti-CXCR4 antibodies were obtained from Santa Cruz biotechnology (Santa Cruz, CA). Rabbit polyclonal anti-Dock180 (C4C12) antibodies were from Cell Signaling Technology (Beverly, MA). Mouse monoclonal anti-RhoG (1F3 B3 E5) was purchased from Millipore (Billerica, MA). Mouse monoclonal anti-Rac1 was from Cytoskeleton (Denver, CO). Mouse monoclonal anti-GFP was purchased from Roche Applied Science (Penzberg, Germany). Mouse monoclonal anti-β-actin antibody was from Sigma Aldrich (St. Louis, MO). Horseradish peroxidase-conjugated anti-mouse, anti-rabbit, and anti-goat secondary antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA). Recombinant Human/Rhesus Macaque/Feline CXCL12/SDF-1α and fibronectin were purchased from R&D systems (Minneapolis, MN)
3
Fluorescent Tagging of DDK-tagged Proteins
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DPCs were seeded and fixed on cell slides. All the slides were blocked in 3% BSA for 30 minutes at room temperature. Mouse monoclonal anti-DDK antibody (1 : 4000, Origene, MD, USA) was used for detection of flag tag. Alexa Fluor® 647 conjugated anti-mouse IgG fragments (1 : 1000, Cell Signaling Technology, MA, USA) staining was performed for 1 h at room temperature. The stained cells were observed under the confocal laser scanning microscopy (Carl Zeiss, Germany).
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