The synthesis of NCT-58 has been described previously [29 (link)]. Primary antibodies targeted Ki-67, CD31, ALDH1 and CD44 (Abcam, MA); HER2, phospho-HER2 (Tyr1221/1222), HER3, phospho-HER3 (Tyr1289), EGFR, phospho-EGFR, Akt, phospho-Akt (Ser473), PARP, cleaved-PARP, cleaved-caspase-3, cleaved-caspase-7, vimentin, Nanog, Oct4, Sox2, Ras, Raf (Ser338), phospho-Raf, Mek, phospho-Mek (Ser217/221), Erk and phospho-Erk (Tyr202/204) (Cell Signaling, CA); CB11 (Thermo Fisher Scientific Fremont, CA); anti-intracellular domain (ICD) HER2 clone 4B5 (Ventana Medical Systems, AZ); survivin, HSP70, HSP90, and HSF-1 (Santa Cruz Biotechnology, CA); and GAPDH (Invitrogen, CA). Secondary antibodies were HRP-conjugated anti-rabbit and mouse IgG (Bio-Rad Laboratories, CA) and Alexa Fluor-488 and −594 goat anti-rabbit IgG (Invitrogen).
Alexa fluor 488 and 594 goat anti rabbit igg
Manufactured by Thermo Fisher Scientific
Alexa Fluor-488 and −594 goat anti-rabbit IgG are fluorescently-labeled secondary antibodies used for detecting and visualizing rabbit primary antibodies in various immunoassays and imaging applications. Alexa Fluor-488 emits green fluorescence, while Alexa Fluor-594 emits red fluorescence when excited by appropriate wavelengths of light.
Automatically generated - may contain errors
Lab products found in correlation
Survivin, by Santa Cruz Biotechnology (2 mentions)
Hsp70, by Santa Cruz Biotechnology (2 mentions)
Gapdh, by Thermo Fisher Scientific (2 mentions)
Phospho egfr, by Cell Signaling Technology (2 mentions)
Phospho her3, by Cell Signaling Technology (2 mentions)
Anti intracellular domain icd her2 clone 4b5, by Roche (2 mentions)
Hrp conjugated anti rabbit and mouse igg, by Bio-Rad (2 mentions)
Cleaved parp, by Cell Signaling Technology (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Phospho mek, by Cell Signaling Technology (1 mentions)
Phospho raf, by Cell Signaling Technology (1 mentions)
Aldh1, by Abcam (1 mentions)
Cleaved caspase 7, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Tanespimycin, by Selleck Chemicals (1 mentions)
Paclitaxel, by Selleck Chemicals (1 mentions)
Hsp27, by Santa Cruz Biotechnology (1 mentions)
Phospho akt, by Abcam (1 mentions)
Aldh1a1, by Abcam (1 mentions)
Phospho mtor, by Abcam (1 mentions)
3 protocols using alexa fluor 488 and 594 goat anti rabbit igg
1
Immunohistochemical Profiling of Cancer Markers
2
Synthesis and Characterization of HVH-2930
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The synthesis of HVH-2930 is described in the Supplementary Information . Tanespimycin, onalespib and paclitaxel were purchased from Selleckchem (Houston, TX). Dimethyl sulfoxide (DMSO), propidium iodide (PI), MG132, N-Acetyl cysteine (NAC) and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO). Trastuzumab was purchased from Roche Diagnostics Korea Co., Ltd (Seoul, South Korea). Primary antibodies used for immunoblotting and immunostaining were obtained as follows: survivin, HSP90, HSP70, HSF1, HSP27, Cyclin D1 (Santa Cruz, CA); HER2 (CD11), Ki-67, Bcl-2, ALDH1A1, CD44, MDR1 and CD31 (Abcam, MA); AKT, phospho-AKT (S473), mTOR, phospho-mTOR (S2448), Bax, PARP, cleaved-PARP, cleaved-caspase-3, cleaved-caspase-7, cleaved-caspase-8, caspase-9, EGFR, phospho-EGFR (Y1068), HER2, phospho-HER2 (Y1221/1222), HER3, phospho-HER3 (Y1289), MEK1/2, phospho-MEK1/2 (S217/221), ERK1/2, phospho-ERK1/2 (T202/Y204), Oct4 and Nanog (Cell Signaling, CA); p27 (BD bioscience, NJ); phospho-HSF1 (S326) (Bioss, MA); anti-intracellular domain (ICD) HER2 clone 4B5 (Ventana Medical Systems, AZ); and GAPDH (Invitrogen, CA). Secondary antibodies were HRP-conjugated anti-rabbit and mouse IgG (Bio-Rad Laboratories, CA) and Alexa Fluor-488 and -594 goat anti-rabbit IgG (Invitrogen).
3
Immunofluorescence Staining of FLAG and TET1
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Cells were fixed with 4% formaldehyde for 15 min at RT and then permeabilized in 0.1% Triton X-100 for 15 min at RT. Cells were then washed by phosphate buffered saline (PBS) for three times, followed by incubation with 5% normal goat serum for 30 min at RT. Subsequently, cells were incubated with primary antibody, the anti-FLAG antibody (Sigma) and anti-TET1 (Genetex), for 2 h at RT. After washing three times with PBS, cells were incubated with secondary antibody, Alexa Fluor 488 and 594 goat anti-rabbit IgG (Invitrogen), for 1h at RT. Finally, cells were washed three times with PBS and mounted using Prolong Gold Antifade Reagent (Invitrogen).
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