MALDI-FTICR mass spectrometric analysis was performed with a Bruker solariX mass spectrometer equipped with a 9.4 T superconducting magnet and SmartBeam™ laser optics. Mass calibrations were performed externally using DHB and a peptide mixture (angiotensin II, substance P, bombesin, and ACTH clip 1–17) (Sigma, St. Louis, MO, USA) as mass standards. External ion accumulation was used in both positive and negative ion mode over a mass range of 200–1800 m/z with a resolution of 200,000 at m/z 200. SolariX Control software was used for data acquisition. For MALDI MSI analysis, mass spectra were acquired across the entire sample section with a SmartBeam II laser operating at 1000 Hz, a laser focus of 25 μm, 1 scan acquired from each matrix spot, and a raster step size of 100 μm. The device parameters for MALDI MSI were chosen as follows: plate offset voltage, 100 V; deflector plate voltage, 180 V. Data were processed using DateAnalysis 4.0 (Bruker Daltonics) and FlexImaging 3.0 software (Bruker Daltonics).
Bombesin
Manufactured by Merck Group
Sourced in United States
Bombesin is a laboratory product manufactured by Merck Group. It is a peptide that functions as a regulatory neuropeptide. The core function of Bombesin is to act as a stimulant for the release of various hormones and neurotransmitters in the body.
Automatically generated - may contain errors
Lab products found in correlation
Angiotensin 2, by Merck Group (1 mentions)
Substance p, by Merck Group (1 mentions)
Fleximaging 3, by Bruker (1 mentions)
Solarix mass spectrometer, by Bruker (1 mentions)
Hs014, by Bio-Techne (1 mentions)
Human α msh, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Cilengitide, by Selleck Chemicals (1 mentions)
Blebbistatin, by Merck Group (1 mentions)
Sb202190, by Bio-Techne (1 mentions)
Lm609, by Merck Group (1 mentions)
Recombinant human tgf β1, by R&D Systems (1 mentions)
Actin antibody, by Merck Group (1 mentions)
Heregulin, by Genentech (1 mentions)
Anti flag affinity gel, by Merck Group (1 mentions)
U0126, by Cell Signaling Technology (1 mentions)
Epidermal growth factor (egf), by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
293t cell, by American Type Culture Collection (1 mentions)
Lncap cell, by American Type Culture Collection (1 mentions)
5 protocols using bombesin
1
MALDI-FTICR Mass Spectrometry for Imaging
2
Melanocortin Receptor Agonist Assay
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Nle(4),D-Phe(7)]α-melanocyte-stimulating hormone (NDP-α-MSH) was from Phoenix Pharmaceuticals, Inc. (Cat. no. 043-06, Burlingame, CA, USA). HS014 was from Tocris (Cat. no. 1831, Abingdon, UK). Human α-MSH, D-glucose and bombesin were from Sigma Aldrich (Cat. no. M4135, G8270 and B4272, Brøndby, Denmark).
3
Radiolabeled Bombesin Analogs Uptake
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PC-3 and HaCaT cells were trypsinized, centrifuged, suspended, and aliquoted in test tubes at a cell concentration of 1 × 106/1 mL RPMI-1640 solution. Tubes were incubated for 60 or 120 min in the presence of 0.37 MBq of [68Ga]Ga-NODAGA-AMBA or [44Sc]Sc-NODAGA-AMBA at 37 °C. In blocking experiments, 200 nM bombesin (Sigma-Aldrich) was added to the cells. After the incubation time, samples were washed three times with ice-cold PBS and the radioactivity was measured with a calibrated gamma counter (Perkin-Elmer Packard Cobra, Waltham, MA, USA) for 1 min within the 68Ga- and 44Sc-sensitive energy window. Decay-corrected radiotracer uptake was expressed as counts/(min × (106 cells)) (cpm). The uptake of the radiopharmaceuticals was expressed as percentage of the total radioactivity of radiotracers added to the cells (%ID/million cells). Each experiment was performed in triplicate and the displayed data represent the means of at least three independent experiments (±SD).
4
Integrin and TGF-β Signaling Modulation
5
Signaling Pathways in Prostate and Kidney Cells
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LNCaP cells and 293 T cells were obtained from the American Type Culture Collection (Manassas, VA, USA). EGF (R&D Systems, Minneapolis, MN, USA), IL-6 (R&D Systems), Gas6 (R&D Systems) and bombesin (Sigma-Aldrich, St Louis, MO, USA), U0126 (Cell signaling, Beverly, MA, USA) were purchased. Heregulin was a gift from Genentech (South San Francisco, CA, USA). Dasatinib was a gift from Bristol-Myers-Squibb (Princeton, NJ, USA). A mouse monoclonal antibody against AR (F39.4.1, Biogenex, San Ramon, CA, USA) was used for immunoblotting and a polyclonal antibody against AR (C-19, Santa Cruz) was used for immunoprecipitation. The antibody against total Ack1 was described previously34 (link). A phospho-specific antibody against Ack1 p-Tyr-284 (#09–142) was obtained from Millipore (Billerica, MA, USA). Antibody against SLIRP (#ab51523) was purchased from Abcam (Cambridge, MA, USA). Antibodies against total ERK (#9102) and phospho-ERK (#9101) were obtained from Cell Signaling Technology (Beverly, MA, USA). Actin antibody (#A3853) and anti-Flag affinity gel (#A2220) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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