Difference between revisions of "Bradford Assay"

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m (formatting change for PMID)
m (Protocol: Added Protein Lysate)
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==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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##Blank then measure samples, absorbance must be less than 0.9
 
##Blank then measure samples, absorbance must be less than 0.9
 
##Print (hit Recall, then enter, then 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 Reagent, 10ul Sample)
 +
#Add 5ul of sample to 100ul of reagent in well
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#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
 
*PMID 942051

Revision as of 21:23, 26 July 2013

Materials

  • BioRad Protein Assay Dye Reagent Concentrate cat#500-0006
  • Disposable Plastic Cuvette

Protocol

Cuvette Bradford Assay

  1. Dilute reagent 5X in water, stable for 2-3 weeks
  2. Pipet 1 mL into disposable plastic cuvette
  3. Add 1-10 uL of protein sample, cover with parafilm and mix
  4. Let sit 5-10 min to react
  5. Set spectrophotometer as follows:
    1. Go to protein assay then Bradford assay
    2. Set formula, then select more
    3. Set b=0.045 (or determine slope)
    4. Set dilution to be 1/vol (ie 0.1 for 10 uL)
    5. Blank then measure samples, absorbance must be less than 0.9
    6. Print (hit Recall, then enter, then print) and attach to experiment

Protein Lysate Bradford Assay

  1. Dilute reagent 5X in water, stable for 2-3 weeks
  2. In a 96 well plate, dilute sample 20X (190ul Reagent, 10ul Sample)
  3. Add 5ul of sample to 100ul of reagent in well
  4. Run "Bradford Assay Protocol" on Plate Reader

Reference