Author Topic: Bradford assay. Please check if I am doing it right way  (Read 6677 times)


  • Newbie
  • *
  • Posts: 1
Bradford assay. Please check if I am doing it right way
« on: July 28, 2013, 11:43:48 AM »
Hi all
     I extracted plant protein using phenol extraction method by methanol precipitation. Could you please check if I am doing bradford assay standard curve and also checking protein concentration from unknown sample.

1) BSA Stock 1mg/ml dissolved in deionised water 2) Bradford reagent from Sigma

I took 5µl of stock BSA and add 995µl of Bradford reagent and checked 0.D at 595 nm and also I took 10µl, 25µl, 30µl, 40µl, 50µl, 60µl, 70µl of stock BSA and add 990, 975, 970, 960, 950, 940, 930 µl of Bradford reagent and did O.D at 595 nm.

BSA (1mg/ml)    Bradford reagent    0.D 595
5 µl                     995 µl             0.106
10 µl                     990 µl             0.236
25 µl                     975 µl             0.544
30 µl                     970 µl             0.690
40 µl                     960 µl             0.791
50 µl                     950 µl             0.861
60 µl                     940 µl             0.882
70 µl                     930 µl             0.911

The value I got from the graph was

y = 16.04x

R² = 0.7773

2) Now I have my protein sample (unknown concentration), I did in the following way.

Protein Sample 1    Bradford    0.D 595
10µl                    990 µl    0.117
40 µl                    960 µl    0.726
50 µl                   940 µl    0.813

Protein Sample 2    Bradford    O.D 595
10 µl                    990 µl    0.135
40 µl                    960 µl    0.545
50 µl                    940 µl    0.821

Protein Sample 3    Bradford    0.D 595
10 µl                   990 µl    0.05
40 µl                   960 µl    0.321
50 µl                   940 µl    0.671

Then I did calculation in this way: Y = 16.04 x   
x= y/16.04 and I got 0.958 mg/ml for sample 1, for sample 2 I got 0.904 mg/ml and for sample 3 I got 0.516 mg/ml

Now I need up to 350 µl for gels, but I need same concentration, so I took the lowest one which is sample 3 (516 µg in 1000 µl) so in 350 µl I got 180.6 µg.

Then I calculated for sample 1 (958µg in 1000 µl) so I got 180 µl and for sample 2 (904 µg in 1000 µl), so I got 199.7 µl

Is this the right way I did to check the known concentration and required concentration? Please help

Thank you


  • Newbie
  • *
  • Posts: 4
    • Buy Replica Christian Louboutin Alti Gold Spikes 160mm Cork Pumps Natural Gold
Buy Replica Christian Louboutin Alti Gold Spikes 160mm Cork Pumps Natural Gold
« Reply #1 on: September 29, 2013, 01:08:27 PM »

Shoes Christian Louboutin Alti Gold Spikes 160mm Cork Pumps Natural Gold, price $156.00.
If you likes  Christian Louboutin Alti Gold Spikes 160mm Cork Pumps Natural Gold, click the "I want to buy" below to find out where it is most advantageous to buy shoes brand Best Fake Christian Louboutin

Category: Christian Louboutin Platforms
Brand: Replicas Christian Louboutin

Tags product: Christian Louboutin Alti Gold Spikes 160mm Cork Pumps Natural Gold, Christian Louboutin, Christian Louboutin Platforms, replica christian louboutin, christian louboutin replica, christian louboutin outlet, christian louboutin sale, christian louboutin shoes outlet, replica christian louboutin shoes, cheap louboutin outlet, christian louboutin fake, christian louboutin shoes sale, cheap christian louboutin shoes sale, cheap louboutin sale, christian louboutin knockoff, christian louboutin, louboutin shoes outlet, christian louboutin outlet online, louboutin, louboutin or, christianlouboutin, shoes christian louboutin, shoes by christian louboutin, shoes from christian louboutin, where can i get christian louboutin shoes, outlet louboutin, christian louboutin red shoes, red christian louboutin shoes, fake christian louboutins, christian louboutins fake, red louboutin
Hello users of  I would like to introduce you to our new web shop, cheap replica brands direct from China.
Purchase goods direct from China - offering multiple products, also offering delivery services purchased anywhere in the world.  For retail customers  - it is an opportunity to buy cheap goods from China, we have created a catalog of extensive directory of links to our stores on this page.  If you are interested in wholesale supply of Chinese products, the managers can assist with finding a suitable product, direct from the factory and ordered to your doorstep.
An important factor is that - it's not just the site of goods from china, the company is headquarted in Guangzhou.  Wholesale clients have a unique opportunity to go directly to the producers, without the middle men involved.
Own office in china, this guarantees you the security of the transactions, and our guarantees applied to borth wholesale and retail customers.
Sometimes, potential buyers do not know where or how to find a product that meets one or another specific needs.  In such situations, you can contact our managers and we shall find the product which you are looking for.
We will find you any pair of shoes of any brand!
On this page we will provide one of the samples of our products
Our other websites:
-christian louboutin red shoes
-outlet louboutin
-cheap christian louboutin shoes sale
-cheap christian louboutin shoes sale
mans shoes
Promo-code: #jhiiiiljkh-99999-hndfdf