Doxercalciferol

Manufactured by Merck Group

Doxercalciferol is a synthetic vitamin D analog used in the manufacture of pharmaceutical and research laboratory equipment. It is a key component in the production of various diagnostic and analytical tools employed in scientific investigations.

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Lab products found in correlation

8 protocols using doxercalciferol

Sorafenib and Autophagy Modulation in Cancer

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Sorafenib (Sf) (Selleckchem, Houston, TX) was used at different concentrations at micromolar scale. Doxercalciferol (1-hydroxyvitamin D2; D2) (Sigma-Aldrich, St. Louis, MO) was used at the final concentration of 100 nM. Carnosic acid (CA) (Enzo Life Sciences Inc., Farmingdale, NY) was used at the final concentration of 10 μM (7 (link),27 (link)). Autophagy inhibitor chloroquine (Sigma) was used at 10 μM and caspase 3 inhibitor Z-DEVD-FMK (Selleckchem) at final concentration of 1 μM. The following antibodies were used for Western blotting: Beclin1 (#3738), Cleaved Caspase 3 (#9661), Capase 9 (#9502), Atg3 (#3415), p62 (Cat#39749S), LC3I/II (#12741) and HRP-linked anti-rabbit (#7074) antibodies were purchased from Cell Signaling Technologies (Danvers, MA). The loading control for Western blotting β-actin antibody was purchased from Sigma-Aldrich. Propidium Iodide Nucleic Acid Stain kit was purchased from Invitrogen (Carlsbad, CA).

Wu Q., Wang X., Pham K., Luna A., Studzinski G.P, & Liu C. (2019). Enhancement of sorafenib-mediated death of Hepatocellular carcinoma cells by Carnosic acid and Vitamin D2 analog combination.. The Journal of steroid biochemistry and molecular biology, 197, 105524.

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Carnosic Acid Regulates Cell Apoptosis

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Carnosic acid (CA) was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY). Arabinocytosine (Cytarbine, AraC) and Doxercalciferol (D2) were purchased from Sigma-Aldrich (St. Louis, MO). ASK1 inhibitor NQDI-1 was from Selleck Chemical Inc (Houston, TX). The antibodies that used for Western blotting as following: TXNIP (#14715), Phospho-ASK1 (Thr845, #3765), ASK1 (#8662), Phospho-MKK4 (Ser 257, #4514), MKK4 (#9152), Phospho-JNK (Thr183/Tyr185, #9255), JNK (#9252), Phospho-p38 MAPK (Thr180/Tyr182, #4511), Phospho-MEK1 (Ser217/221, #9121), MEK1 (#9122), cleaved caspase 9 (#9505), cleaved caspase 3 (#9661) and HRP-linked antirabbit (#7074) antibodies were purchased from Cell Signaling Technologies. BIM (sc-8625), VDR (sc-1008), p38 (sc-448) and CRK-L (sc-319) were obtained from Santa Cruz Biotechnology (Dallas, TX).

Wang X., Nachliely M., Harrison J., Danilenko M, & Studzinski G. (2018). PARTICIPATION OF VITAMIN D-UPREGULATED PROTEIN 1 (TXNIP)-ASK1-JNK1 SIGNALOSOME IN THE ENHANCEMENT OF AML CELL DEATH BY A POST-CYTOTOXIC DIFFERENTIATION REGIMEN.. The Journal of steroid biochemistry and molecular biology, 187, 166-173.

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AML Cell Line Cytotoxicity Assay

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Two AML cell lines, HL60 (cultured from a patient with acute promyeloblastic leukemia [24 (link)], and U937 (monocytes from histiocytic lymphoma) [25 (link)], were cultured as previously described [21 ].
Arabinocytosine (AraC) and Doxercalciferol (1α-hydroxyvitamin D2; D2) were purchased from Sigma-Aldrich (St. Louis, MO). Carnosic acid (CA) was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY). BRAF inhibitors SB590885 and TAK632 were purchased from Selleck Inc., stated to inhibit both wild type and mutated BRAF. The antibodies used for Western blots were: BRAF, CRAF, BIM (sc-8625), VDR (sc-1008), and CRK-L, the V-Crk Avian Sarcoma Virus CT10 Oncogene Homolog-Like (sc-319), were obtained from Santa Cruz Biotechnology (Dallas, TX). Phospho-H2AX-Ser139 (#9718), phospho-BIM-Ser69 (#4581) and HRP-linked anti-rabbit (#7074) antibodies were from Cell Signaling Technologies (Danvers, MA). The siRNA transfection reagents were from Santa Cruz Biotechnology (Dallas, TX).

Wang X., Harrison J.S, & Studzinski G.P. (2016). BRAF signals to pro-apoptotic BIM to enhance AraC cytotoxicity induced in AML cells by Vitamin D-based differentiation agents. The Journal of steroid biochemistry and molecular biology, 173, 139-147.

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Apoptotic Pathways Activation Assay

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Arabinocytosine (AraC) and Doxercalciferol (1α-hydroxyvitamin D₂; 1-D2) were purchased from Sigma-Aldrich (St. Louis, MO). Carnosic acid (CA) was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY). JNK inhibitor SP600125 was from Cell Signaling Inc (Danvers, MA). The antibodies that used for Western blotting as following: BRAF (#9433), CRAF (#9422), Phospho-JNK (Thr183/Tyr185, #9255), JNK (#9252), Phospho-p38 MAPK (Thr180/Tyr182, #4511), Phospho-C-JUN (Ser63, #2631), C-JUN (#9165), Phospho-BIM (Ser69, #4585), Phospho-H2AX (Ser139, #9718), FOXO3a (#2497), cleaved Caspase 9 (#9505), cleaved caspase 3 (#9661) and HRP-linked anti-rabbit (#7074) antibodies were purchased from Cell Signaling Technologies. BIM (sc-8625), VDR (sc-1008) and Crk-L (sc-319) were obtained from Santa Cruz Biotechnology (Dallas, TX).

Wang X., Beute W.K., Harrison J.S, & Studzinski G.P. (2017). JNK1 AS A SIGNALING NODE IN VDR-BRAF INDUCTION OF CELL DEATH IN AML. The Journal of steroid biochemistry and molecular biology, 177, 149-154.

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Molecular Mechanisms of Carnosic Acid

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Arabinocytosine and Doxercalciferol were purchased from Sigma-Aldrich (St. Louis, MO). Carnosic acid (CA) was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY). The following antibodies: Crk-L (sc-319), VDR (sc-1008), C/EBP beta (sc-150), and p27Kip1 (sc-528) were obtained from Santa Cruz Biotechnology (Dallas, TX). Phospho-Ser139-H2AX (#9718), phospho-Chk1 (#2348), and HRP-linked anti-rabbit (#7074) antibodies were purchased from Cell Signaling Technologies (Danvers, MA).

Wang X., Harrison J.S, & Studzinski G.P. (2015). Enhancement of Arabinocytosine (AraC) toxicity to AML cells by a differentiation agent combination. The Journal of steroid biochemistry and molecular biology, 164, 72-78.

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Caspase Inhibitors Enhance AraC Cytotoxicity

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Arabinocytosine and Doxercalciferol (1α-hydroxyvitamin D₂; 1-D2) were purchased from Sigma-Aldrich (St. Louis, MO). Carnosic acid (CA) was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY). The antibodies used for Western blots were: Bim (sc-8625), VDR (sc-1008), C/EBP beta (sc-150) and Crk-L (sc-319) were obtained from Santa Cruz Biotechnology (Dallas, TX). Phospho-Ser139-H2AX (#9718), phospho-Ser345-Chk1 (#2348), caspase 3, caspase 9 and HRP-linked anti-rabbit (#7074) antibodies were from Cell Signaling Technologies (Danvers, MA). The siRNA transfection reagents were from Santa Cruz Biotechnology (Dallas, TX), and Caspase 3 inhibitor (Z-DEVD-FMK) from Santa Cruz Biotechnology. Specific caspase 1 inhibitor (Z-WEHD-FMK) and negative control for caspase inhibitors (Z-FA-FMK) were from Abcam (Cambridge, MA). AML cells were pretreated with 100 nM AraC for 72 h, then incubated with 10 μM of caspase inhibitors for 1 h before adding other agents.

Harrison J.S., Wang X, & Studzinski G.P. (2016). The role of VDR and BIM in potentiation of cytarabine–induced cell death in human AML blasts. Oncotarget, 7(24), 36447-36460.

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Vitamin D Derivatives and Kinase Inhibitors Protocol

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1,25D was a kind gift from Dr. Milan Uskokovic (Bioxell). Doxercalciferol (1α-hydroxyvitamin D₂; 1-D₂) was purchased from Sigma-Aldrich (St. Louis, MO). The following antibodies: P-MEK5 (Ser311/Thr315, sc-135702), MEK5 (sc-10795), P-MEF2C (Thr300, SC-130201), and Crk-L (sc-319) were obtained from Santa Cruz Biotechnology (Dallas, TX). P-C/EBPβ (Thr235, #3084), P-C/EBPα (Thr222/226, #2844), MEF2C (#5030), ERK5 (#3372), P-ERK5 (Thr187/Tyr220, #3371), P-ERK1/2 (Thr202/Tyr204, #4370), anti-rabbit (#7074) and anti-mouse (#7076) secondary antibodies conjugated to HRP were purchased from Cell Signaling Technologies (Danvers, MA). The pharmacological inhibitors of ERK5 kinase MEK5 (BIX02189), and ERK5 autophosphorylation (XMD8-92) were purchased from Selleck Chemicals (Houston, TX) and Santa Cruz Biotechnology Inc., respectively. The MEK1/2 inhibitor PD98059 was obtained from Cell Signaling Technologies. Nitrocellulose membranes were purchased from GE Healthcare (Pittsburgh, PA). The vitamin D derivatives (VDDs) were dissolved in ethanol and kinase inhibitors in DMSO.

Wang X., Pesakhov S., Harrison J.S., Danilenko M, & Studzinski G.P. (2014). ERK5 Pathway Regulates Transcription Factors Important for Monocytic Differentiation of Human Myeloid Leukemia Cells. Journal of cellular physiology, 229(7), 856-867.

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Doxercalciferol Enhances Regeneration

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A Vitamin D2 analogue, Doxercalciferol (Sigma Aldrich), was re-suspended in DMSO to a concentration of 10g/L and subsequently diluted in water. Early stage blastemas, generated as a result of amputation, were treated with one of two concentrations of the analogue, 1 mg/kg and 10 mg/kg of body weight by injecting anesthetized animals intraperitoneally in the flank with the analogue.

Vieira W.A., Wells K.M., Milgrom R, & McCusker C.D. (2018). Exogenous Vitamin D signaling alters skeletal patterning, differentiation, and tissue integration during limb regeneration in the axolotl.. Mechanisms of development, 153, 1-9.

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