@@thamizh2850 What do you mean by linear range? So the concentration of protein in the soy milk is enough for the Bradford Assay and spectrophotometer to work?
what kind of non covalent bonding the BSA binds to protein?can you explain what happens during the incubation time,could you explain the mechanism?why does it cause a shift in maximal absorbance?
vanderwalls and ionic bonding is taking place, the bradford reagent interacts with the protein linearly. such that more protein in the sample results in more non-covalen bonding between the dye and the protein. for more detailed info on the principle behind this mechanism see en.wikipedia.org/wiki/Bradford_protein_assay
basically, you can't directly quantify the amount of protein in an unknown sample. You need to do a relative quantification - which means by using a series of known protein concentration, you obtain a set of absorbance values which are arranged to form a standard curve. Then, the standard curve is used to indirectly quantify the amount of protein you have in your unknown sample based on its absorbance values.
Is the Phosphate buffered saline a mandatory buffer to use, i don't have that buffer. If i don't use it what will happen to the pH, and is their any other buffer i can use.
For a coloured protein binding sample such as polysaccharide of inonotus obliquus, which is usually brown in colour. How should I approach it since it is not white in colour such as the milk.
make a 10x - 20x dilution or something, quantify it, before multiplying back by the dilution factor. Once you do the dilution, the colour should be less prominent.
What is the procedure for qualitative estimation of proteins by Bradford assay?
Will this work if I want to test protein concentration in soy milk?
@@thamizh2850 Thank you
@@thamizh2850 What do you mean by linear range? So the concentration of protein in the soy milk is enough for the Bradford Assay and spectrophotometer to work?
Can we have a sprout seed with protein concentration of 55 mg/ml?
@@thamizh2850 will this method work for the determination of protein in food such as rice or wheat?
@@thamizh2850 thanks
Does it matter if I use a 3.5 ml cuvette? Will it affect the absorbance?
Need to read: "Lambert-Beer law" in order to understand the correlation between absorbance, concentration and dilution factor.
i want to estimate the protein of pollen, does anyone have a procedure or info to help??
what kind of non covalent bonding the BSA binds to protein?can you explain what happens during the incubation time,could you explain the mechanism?why does it cause a shift in maximal absorbance?
vanderwalls and ionic bonding is taking place, the bradford reagent interacts with the protein linearly. such that more protein in the sample results in more non-covalen bonding between the dye and the protein. for more detailed info on the principle behind this mechanism see en.wikipedia.org/wiki/Bradford_protein_assay
@@alisaguadarroma829 thankyou so much!!
What is the purpose of standard,I couldn’t get it,please explain to me!?
basically, you can't directly quantify the amount of protein in an unknown sample. You need to do a relative quantification - which means by using a series of known protein concentration, you obtain a set of absorbance values which are arranged to form a standard curve. Then, the standard curve is used to indirectly quantify the amount of protein you have in your unknown sample based on its absorbance values.
does it work for chicken?
Is the Phosphate buffered saline a mandatory buffer to use, i don't have that buffer. If i don't use it what will happen to the pH, and is their any other buffer i can use.
It just matters that you use the same buffer as your sample is in.
So your control is just the buffer.
For a coloured protein binding sample such as polysaccharide of inonotus obliquus, which is usually brown in colour. How should I approach it since it is not white in colour such as the milk.
make a 10x - 20x dilution or something, quantify it, before multiplying back by the dilution factor. Once you do the dilution, the colour should be less prominent.
I want to estimate the protein from the fish sample.. Can u suggest for me procedure plzzz...
yes i can suggest u procedure
thank you for having such video it was helpful
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