changes in protein profiles in bortezomib applied multiple myeloma ...

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B.3.1. XTT Reaction Solution In order to prepare a reaction solution sufficient for a 96-well plate, XTT reagent was defrosted and the Activation reagent was immediately defrosted prior to use in a 37 o C bath. Then 0,1 ml Activation reagent was added to 5 ml XTT reagent. B.4. Caspase-3 Colorimetric Assay Kit Caspase-3 colorimetric assay kit was obtained from BioVision, (USA). B.4.1. 2X Reaction Solution DTT was added to the 2X Reaction Buffer immediately before use. 10 mM final concentration :Add 10 µl of 1.0 M DTT stock per 1 ml of 2X Reaction Buffer. Table B.2. Preparation of 2X Reaction Buffer 2X Reaction Solution ml 1.0 2X Reaction Buffer µl 990 Dithiothreitol DTT 0,6 594 6.0 B.4.2. Stock BSA Standard (1 mg/ml) 0.01 g of BSA was dissolved in 10 ml PBS. µl 10 105
B.4.3. Standard Curve for BSA Table B.3. Absorbance Values for BSA Standards Concentrations (µg/µl) Absorbances (595 nm) 1 0,1775 2 0,182 4 0,191 8 0,209 20 0,263 40 0,353 Figure B.3. Standard Curve for BSA Figure B.1. Standard Curve for BSA B.4.4. Coomassie Brilliant Blue G-250 (CBB G-250) Dye 100 mg Coomassie Brilliant Blue G-250 (CBB G-250) 50 ml Absolute Ethanol 100 ml 85% Phosphoric Acid The above-mentioned mass of CBB G-250 was weighed and dissolved in 50 ml of absolute ethanol. Then, 100 ml of 85% Phosphoric Acid was added to this solution. After the overall solution was filtered through filter paper, the filtered solution (~15-20 ml) was diluted to 1 liter with ultrapure water and stored at 4 o C. y = 0.1730 + 0.0045x R 2 = 0.9966 106
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CHANGES IN PROTEIN PROFILES IN BORT
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ACKNOWLEDGEMENTS First of all, with
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ÖZET BORTEZOMĠB UYGULANAN MULTĠP
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2.1.4.1.2. Two-Dimensional Gel Elec
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LIST OF FIGURES Figure Page Figure
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LIST OF TABLES Table Page Table 1.1
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When normal cells in the body begin
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Figure 1.2. Development of Cancer S
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the myeloma cells does not help fig
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A rare form of MM called Nonsecreto
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1.2.3. Risk Factors for MM A risk f
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of the mineralizations by adjusting
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To measure the levels of red cells,
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The levels of blood urea nitrogen (
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By using the system introduced by D
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1.2.7.2. Biology of The Proteasome
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Figure 1.10. Ubiquitin-Proteasome P
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Figure 1.12. Effect of Bortezomib o
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encompasses the investigation of pr
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of modification. Ubiquitin is a sma
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the given cells. On the other hand,
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is-acrylamide (N,N'-methylene-bisac
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2.1.4.2. Detection of Protein Spots
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ange of 600-2500 Da. This is the id
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ions resolved by the mass analyzer
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2.1.4.5.2. Mass Analyzers Figure 2.
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There is no doubt that different ma
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mask low-quantity proteins that may
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3.1. Materials 3.1.1. Medias CHAPTE
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3.2.5. Cell Proliferation Assay Ant
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After 72 hour incubation period at
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eceptors of the Tumor Necrosis Fact
Page 65 and 66: The preparation of stock BSA Standa
Page 67 and 68: 3.2.6.3. Detection of the Loss of M
Page 69 and 70: centrifugation, the supernatant was
Page 71 and 72: selected symmetrically to load the
Page 73 and 74: approximately 35 ml, the volume of
Page 75 and 76: After electrophoresis was finished,
Page 77 and 78: 7 Day Two: Reduction, Alkylation, a
Page 79 and 80: CHAPTER 4 RESULTS AND DISCUSSION 4.
Page 81 and 82: % Changes in Cytoplasmic/Mitochondr
Page 83 and 84: Table 4.1. Magnified Segments of Di
Page 85 and 86: Table 4.1. Magnified Segments of Di
Page 87 and 88: After labeling, black and red color
Page 89 and 90: 78 7 8 Y2S Y3S Table 4.2. Results o
Page 91 and 92: 80 12 S4Ca Table 4.2. Results of Di
Page 93 and 94: 82 27 Y5S Table 4.2. Results of Dif
Page 95 and 96: Spot 4, Ubiquitin-conjugating enzym
Page 97 and 98: is a transcriptional repressor, but
Page 99 and 100: activity and loss of mitochondrial
Page 101 and 102: distributed, but correlate with nat
Page 103 and 104: DeMartino, G. N. and Slaughter, C.
Page 105 and 106: proteomic analyses of a systematica
Page 107 and 108: Mahindraa, A., Hideshimab, T. and A
Page 109 and 110: Richardson, P. G., Hideshima, T. an
Page 111 and 112: Terpos, E. 2008. Bortezomib Directl
Page 113 and 114: A.1. RPMI-1640 Growth Medium APPEND
Page 115: Table B.1. Chemicals and Reagents U
Page 119 and 120: Urea (8M) 4.8 g CHAPS (%2) 0.2 g DT
Page 121 and 122: E. F. G. STOCK REAGENT PREPARATION
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