Hello, I rely on your help. On February 27th, I prepared a Bradford reactive, left it for the night at room temperature, filtered it out on the next day and made a calibration graphic with egg albumen. An optical density of samples with different protein content constituted from 0.0237 to 0.0933. Ready Bradford reactive kept in the fridge then.
On 7th March I used this reactive for determination of protein content in Chlorella cells. OD constituted values, that were within ones of the calibration graphic.
Yesterday I did the same, but the values didn't differ and constituted 0.8430, something like that for all samples. I checked OD of plain Bradford reactive and it was just the same.
What might be wrong with the reactive? Do I need to prepare a new one? Or recalibrate the old? And what might such problems be connected with? I've never worked with Bradford reactive and I just don't know how to act.
Please, sorry for grammar mistakes, if there're they. I'm not a native English speaker.
55 minutes ago, Fanipal said:
Hello, I rely on your help. On February 27th, I prepared a Bradford reagent, left it for the night at room temperature, filtered it out on the next day and made a calibration graphic with egg albumen. An optical density of samples with different protein content constituted from 0.0237 to 0.0933. Ready Bradford reagent kept in the fridge then.
On 7th March I used this reagent for determination of protein content in Chlorella cells. OD constituted values, that were within ones of the calibration graphic.
Yesterday I did the same, but the values didn't differ and constituted 0.8430, something like that for all samples. I checked OD of plain Bradford reagent and it was just the same.
What might be wrong with the reagent? Do I need to prepare a new one? Or recalibrate the old? And what might such problems be connected with? I've never worked with Bradford reagent and I just don't know how to act.
Please, sorry for grammar mistakes, if there're they. I'm not a native English speaker.
57 minutes ago, Fanipal said:
Hello, I rely on your help. On February 27th, I prepared a Bradford reactive, left it for the night at room temperature, filtered it out on the next day and made a calibration graphic with egg albumen. An optical density of samples with different protein content constituted from 0.0237 to 0.0933. Ready Bradford reactive kept in the fridge then.
On 7th March I used this reactive for determination of protein content in Chlorella cells. OD constituted values, that were within ones of the calibration graphic.
Yesterday I did the same, but the values didn't differ and constituted 0.8430, something like that for all samples. I checked OD of plain Bradford reactive and it was just the same.
What might be wrong with the reactive? Do I need to prepare a new one? Or recalibrate the old? And what might such problems be connected with? I've never worked with Bradford reactive and I just don't know how to act.
Please, sorry for grammar mistakes, if there're they. I'm not a native English speaker.
Yes, I just wrote "reactive" instead of "reagent" 😖
We have placed cookies on your device to help make this website better. You can adjust your cookie settings, otherwise we'll assume you're okay to continue.
What's wrong with Bradford?
in Biochemistry and Molecular Biology
Posted
Hello, I rely on your help. On February 27th, I prepared a Bradford reactive, left it for the night at room temperature, filtered it out on the next day and made a calibration graphic with egg albumen. An optical density of samples with different protein content constituted from 0.0237 to 0.0933. Ready Bradford reactive kept in the fridge then.
On 7th March I used this reactive for determination of protein content in Chlorella cells. OD constituted values, that were within ones of the calibration graphic.
Yesterday I did the same, but the values didn't differ and constituted 0.8430, something like that for all samples. I checked OD of plain Bradford reactive and it was just the same.
What might be wrong with the reactive? Do I need to prepare a new one? Or recalibrate the old? And what might such problems be connected with? I've never worked with Bradford reactive and I just don't know how to act.
Please, sorry for grammar mistakes, if there're they. I'm not a native English speaker.
Yes, I just wrote "reactive" instead of "reagent" 😖