Bradford Assay
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- Опубликовано: 7 сен 2024
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This video demonstrates how to quantitate proteins using the Bio-Rad Got Protein?™ kit. Protein concentration is determined both visually by comparing samples to a set of standards and by measuring absorbance on a SmartSpec™ spectrophotometer.
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thank you for having such video it was helpful
What is the procedure for qualitative estimation of proteins by Bradford assay?
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!!
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.
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 of pollen, does anyone have a procedure or info to help??
Will this work if I want to test protein concentration in soy milk?
Yes it should work.
@@thamizh2850 Thank you
Soy milk rich in protein. So make serial dilution to get a linear range.
@@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?
For example, the absorbance value of 2 mg/ml is twice as the absorbance value of 1 mg/ml. But you can not get this if concentration is beyond the saturation limit. I mean no enough reagent component to react with protein in excess and therefore no further increase in absorbance.
does it work for chicken?
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.
I want to estimate the protein from the fish sample.. Can u suggest for me procedure plzzz...
yes i can suggest u procedure
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