Reagents

• 121.1g Tris-Base
• 576.7g Glycine
• 40g SDS
• Bring to 4L DI Water
• Adjust pH to 8.63

• 15g Tris-Base
• 71.25g Glycine
• 10mL 5M EDTA
• Bring to 500mL H₂O
• Adjust to pH 8.5

• 484g Tris
• 200mL 0.5M EDTA, pH 8.0
• 114.2mL Glacial Acetic Acid
• Bring to 2L H₂O

• 134.9g Dulbecco's Modified Eagle's Medium (powder)
• 500mL FBS
• 100ml 100x Streptomycin
• 37g NaHCO₃
• Bring to 10L in ddH₂O
• Filter through 0.2uM sterile filter

• 103.92g RPMI medium 1640 (powder)
• 500mL FBS
• 100mL 100x Streptomycin
• 20g NaHCO₃
• Bring to 10L in ddH₂O
• Filter through 0.2uM sterile filter

• 626mL IMDM
• 313mL Nutrient mixture
• 50mL FBS
• 10mL 100x Pen/Strep
• 500uL 10mg/mL Insulin
• 40uL 1mg/mL Hydrocortisone
• 35uL 100ng/uL EGF
• Fitler through 0.2uM sterile filter

• 80mL FBS
• 20mL DMSO
• Filter through 0.2uM sterile filter

Running Buffer:

• 200mL 10x SDS
• 1800mL H₂O

20x TBS:

• 96.8g Tris-base
• 654g NaCl
• Bring to 4L H₂O
• Adjust to pH 7.4

TBST:

• 200mL 20x TBS
• 4mL Tween 20
• Bring to 4L H₂O

Transfer Buffer:

• 400mL 10x SDS
• 800mL Methanol
• 2800mL H₂O

Stripping Buffer:

• 31.25mL Tris-HCl, pH 6.7
• 10g SDS
• Bring to 500mL H₂O
• Add 390uL BME per 50mL

Blocking Buffer:

• 3g BSA powder
• 100mL TBST
• Vacuum filter through 0.2uM sterile filter

• 1x Protease Inhibitor Cocktail (Sigma)
• 0.5% TX-100
• 50mM b-GP
• 2mM MgCl₂
• 0.1M Na₃VO₄
• 1mM EGTA
• 1mM DTT
• 1x Aprotinin
• 0.1mM PMSF
• 0.3M NaCl
• Adjust pH to 7.2

• 312.5mM Tris
• 25% Glycerol
• Adjust to pH 6.8
• 10% SDS
• 0.005% Bromophenol Blue
• 5% &beta-mercaptoethanol

• 12.5g Luria-Broth powder
• 15g/L Bacto-Agar
• Autoclave 15min
• Cool to 55 degrees C
• 1000x Ampicillin
• Pour into 10cm culture dishes
• Refridgerate once solidified

EGFR Inhibitors	FGFR Inhibitors	Jak/STAT Inhibitors	MAPK Inhibitors
AZD 8931 (pan-ErbB)	AZD 4547	Ruxolitinib	Trametinib
AZD 9291	Ponatinib		Selumetinib
Cetuximab