Bradford Assay: Difference between revisions
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*BioRad Protein Assay Dye Reagent Concentrate cat#500-0006 | *BioRad Protein Assay Dye Reagent Concentrate cat#500-0006 | ||
*Disposable Plastic Cuvette | *Disposable Plastic Cuvette | ||
*0.1mg/mL BSA in H20 as standard | |||
==Protocol== | ==Protocol== | ||
Cuvette Bradford Assay | |||
#Dilute reagent 5X in water, stable for 2-3 weeks | #Dilute reagent 5X in water, stable for 2-3 weeks | ||
#Pipet 1 mL into disposable plastic cuvette | #Pipet 1 mL into disposable plastic cuvette | ||
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##Set dilution to be 1/vol (ie 0.1 for 10 uL) | ##Set dilution to be 1/vol (ie 0.1 for 10 uL) | ||
##Blank then measure samples, absorbance must be less than 0.9 | ##Blank then measure samples, absorbance must be less than 0.9 | ||
##Print and attach to experiment | ##Print (hit Recall, then enter, then print) and attach to experiment | ||
Protein Lysate Bradford Assay | |||
#Dilute reagent 5X in water, stable for 2-3 weeks | |||
#In a 96 well plate, dilute sample 20X (190ul H2O, 10ul Sample)--this dilution factor is tissue-dependent, only need to dilute fat ~5x | |||
#Add 5ul of 20x diluted sample to either 100ul or 200ul of Bradford reagent in well. | |||
##Prepare a standard curve by adding 0.0-3.0ul BSA std (1mg/ml) in increments of 0.5ul for 100 ul or 0-6ul BSA std in increments of 1ul for 200ul. | |||
#Run "Bradford Assay Protocol" on Plate Reader | |||
==Reference== | ==Reference== | ||
Wikipedia: [[wikipedia:Bradford_protein_assay|Bradford Protein Assay]] | *Wikipedia: [[wikipedia:Bradford_protein_assay|Bradford Protein Assay]] | ||
*PMID 942051 | |||